text:
Fear the Walking Dead: Reclamation
Posted by Jon Hicks | Nov 26, 2021 | Fear the Walking Dead, Television
We have a flashback start with Al, she is getting interviewed by someone, which if you piece together the clues, is Isabella. It is obvious that Al is not used to being on that side of the camera. Present day we see Morgan looking for Al, he finds the truck but can't locate Al. So he finds a camera and starts to record a message because he knows she will watch it. Morgan says he heard that she got separated from the others on the way back but he starts to think that she didn't come back on purpose. Al was off on a lead, and heard a transmission from the CRM talking about reclamation, she knows what that means, which is essentially a clear mission so she heads back to the truck.
The soldiers are already close to sweeping the area so she hides under the truck, after the soldiers move on she looks around and sees her camera and tapes are missing and starts to freak out a bit. Morgan comes down from the top of the truck and holds a knife to Al's throat, once they figure out who each other are they chat briefly. Al says that the others shouldn't have told Morgan the information they did about the helicopter and that if they stay here they will die. So Morgan says it's time to move on and also find Grace who is also looking for Al.
Al and Morgan start making their way to the rendezvous point that Morgan and Grace set up, which is about 2 miles away from their current location. Al wanted to wait until night to make the trip but Morgan insisted he was going now. Next scene they were under a road, with a lot of abandoned cars and they say that the meeting spot is at the end of this run of cars. While leisurely walking some stray bullets whiz past them from the soldiers. The two zig zag through the car maze towards the exit. The pair talk while in hiding and Al says the reclamation team is apart of a society and their mission is to eliminate any trace they were ever there, and right now that means them, even though they are not the reason the are here right now.
Grace gets to the meeting point even though Al wasn't sure that she would, Al insists to lead the soldiers away so Grace and Morgan can get away but Morgan says he is not leaving without her, so she relents and gets to the car. Soldiers hit the car with some bullets and maybe the fuel tank so they have to hide the car. The trio catch up a bit about Grace's baby, and how now they are de facto parents of another baby. Grace and Morgan insist to help, so Al comes up with a plan that the MRAP has enough firepower to take them out, and they can get fuel and ammo from the helicopter if she leads them away. So the groups head on their missions, but the helicopter did have gas but no ammunition. Morgan radio's to Al, and she says she knows, it's a light helicopter they don't have ammo, and they have a kid now and they should go back to her. She says she can take it from here, she knows she might die but she wants to take the soldiers with her and refuses to tell them where she is.
Lennie James as Morgan Jones, Maggie Grace as Althea, Karen David as Grace – Fear the Walking Dead _ Season 7, Episode 6 – Photo Credit: Lauren "Lo" Smith/AMC
Al believes the soldiers have her camera and wants it back, but Grace in the truck now knows that it was in Morgan's pack all along. She looks at the tape for clues, but doesn't find anything useful. Al wants to bring the soldiers to a location with a cannon to blow them up, she tests it on a walker at first. The soldiers want to cancel the meetup plans after capturing Morgan and Grace, Al wants to have them let go but the soldiers refuse. The soldiers tell Al they will kill the male, they fire two warning shots and are about to hit the three count when Al tells them she knows where "she" is. She is, "Ground 17", that they are looking for. The soldiers say they require confirmation, so Al says she will show them where she is at, she is at a cabin of her former copilot. After the information the soldiers decide they will kill Morgan and Grace anyway, but the pair are able to wrestle a gun away from a soldier. Morgan holds the second soldier at gunpoint to let Grace go. He also fires off two warning shots, and before the third the soldier lets Grace up. Grace runs to the truck while Morgan wrestles with one of the soldiers, Grace tries to run over the soldier and Morgan uses that to get into the truck. Al tells Grace where to lead the soldiers, which is where Al is.
The soldiers catch up to the truck and end up flying past it in the helicopter to go land and to get to the meeting spot before them. The soldiers do arrive a few seconds before Morgan and Grace, the soldiers shoot some walkers. But Grace pulls up in the truck, Al hops inside and tells Grace that when she says, she needs to just drive. A walker messes up the cannon plans a bit, so Morgan has to go fix it while Al stalls. Al comes out with a tape, the soldiers want the tape, while Al wants the others to be let go, the soldiers refuse so Morgan yells to Grace to "Go Now". Grace moves the truck, Morgan unleashes the cannon, splat goes the two soldiers.
On the trip back, Al tells Morgan this doesn't change anything, she is still going. Morgan says that he saved her life so she owes him an interview. Morgan said if she is really leaving he wants something to remember her by. Al tells Morgan that she knows where the pilot is, but she isn't going to her because she couldn't give up searching for stories so she refused to go with her. Morgan asked enough tough questions that Al had to stop the interview. She heads off after Grace tells her, whoever you think you are letting down wouldn't want you to miss out because of it.
Lennie James as Morgan Jones – Fear the Walking Dead _ Season 7, Episode 6 – Photo Credit: Lauren "Lo" Smith/AMC
Al went back to the site they blew up the soldiers to look for clues. Morgan meets up with her shortly after, Al tells him she made up her mind, and Morgan says you and John tried one more time after I said no and I am doing the same thing here. One of the transmitters starts going off so Al goes back to the helicopter to get some clearer information. The reclamation team is being pulled back to a new target, and Al replies back acting as the team. Ground control says that they have the coordinates of the helicopter and Al says to herself, they will find her no matter where she goes. She replies to ground control that they are in route. Before she goes, Morgan tells her that she is apart of the story, she isn't not apart of things like a piece of machinery like that camera. She is apart of the story, he is glad he was able to be apart of her story and glad that she was able to be apart of his. She says me too, and Morgan tells her to go.
Al makes it to Isabella's cabin, the chimney is putting out smoke so she knows someone is alive. She goes inside, the TV was on, and she plays the tape that was in the VCR. It is of an interview with Al, Isabella comes in and has her at gunpoint. Al tells Isabella that they know where her helicopter is and are probably a couple days behind her, and she wants to go with Isabella. She confirms that they will never stop being chased and will have to continue to move around, and that Al needs to give up chasing a story. To prove it Al breaks her camera right in front of Isabella, she said it was never the camera, it was just that she was scared. But what was scary was not coming here, not trying. Hopefully you are happy with the progress of this storyline, because this is the last we see Al this season.
PreviousAkira Kurosawa: The Honor of Influence
NextNerdy Merchandise for 11/22/21
Jon Hicks
I am a wise cracking avid gamer, drawn more to the role playing games with the vast stories and characters. I collect Funko pops, my name is Riddler3 on the Funatics board. I'm a big DC fan, none bigger than Green Lantern as evidenced by my leg tattoo. My biggest fandom is zombies, namely all things Walking Dead. I love everything about zombies and can talk about it for hours. I enjoy Game of Thrones, sports, basketball, and football at the forefront. I support my hobbies as an office manager at a sign company. Please don't be afraid to reach out and talk about all sorts of topics.
American Horror Story: Roanoke (S6; E9)
Krypton 2×8: "Mercy," Recap & Review
Originals: 8 Spoilers to Know About the Final Season
Will & Grace – Grandpa Jack


Top professionals know that continued education is the key to a successful career.
Defining and detailing a haircut using shears.
Tips and tricks for mastering razors and clippers.
Best practices for working with different curl patterns including the S-Curl to create trendy soft waves.
Enhancing your cut with the perfect color placement.
You will upgrade your skill set and learn to move beyond any fears you have regarding cutting hair.
Complimentary refreshments and wine will be served. Early bird pricing and student discounts are available.


Serving Wilmington, Newark, New Castle, Hockessin, and surrounding areas.
Clean Air Lawn Care is a full-service sustainable and organic lawn care service for residential and small commercial properties in Hockessin, Wilmington, Newark, New Castle and surrounding areas. Local owners, Tim and Ashley are passionately committed to clean practices and exceptional customer support in Delaware with all-natural, chemical-free lawn treatment programs safe for pets and kids and by providing customized service to fit every customers needs.
Help us take a stand against loud, smelly, chemical lawn care. We use all electric equipment that runs on clean, quiet energy from the sun. Check out information about our solar-electric powered mowing, here. Our sustainable mowing and lawn maintenance in North Delaware help to keep neighborhoods quiet, reduce the use of gasoline and oil, and help keep the local communities clean, healthy and beautiful.
Clean Air Lawn Care trucks are mounted with solar panels that allow us to recharge as we work, and allow us to proudly offer emission-free lawn care throughout North Delaware. In addition, our organic fertilization programs are tailored to give you a beautiful lawn in a clean, non-toxic way. Chemical lawn care treatments contaminate waterways and harm your pets or family. Our unique organic lawn fertilization treatment programs include living micro-biology and nutrients to bring your soil and grass to life. Add in a compost topdressing, safe weed control, or a yard clean up to grow your North Delaware lawn the way Mother Nature intended. Get the healthiest lawn around filling out our estimate form today!
You Deserve a Beautiful, Safe, and Natural Lawn!


Prague Attractions | Let's Go for a Trip!
Installed in 1410, the Prague Astronomical clock is the third oldest astronomical clock in the world and the oldest one still working.
When the clock strike every hour from 9am to 9pm, the procession of the Twelve Apostles and other figures on the astronomical clock are set in motion. The apostles will appear in the windows of the clock, where each can be recognised by his attributes. Do keep a close watch on your belongings while watching the clock!
Visitors can go to the top of the Old Town Hall Tower to get a panorama view of Prague.


So we all have heard the clock is "ticking"…but exactly where is this clock and when does the alarm actually go off?–and is there a snooze button!?
Here's what you need to know about age and your fertility on my First Response-Know Sooner Blog. Don't forget to add the website to your favorites for useful tools that can help you keep track of your cycle and fertility.


Artist's Magazine is the leading magazine for serious, active artists. Expert advice on working with pastels, watercolor, oil, acrylics, pen and ink, or any imaginable medium, written by artists for artists.
Be the first person to tell us what you think about Artist's Magazine and you'll save £2.50 on your next purchase!


The Creeps accurately recreates the iconic Warren horror comic magazines popular in the 1970's by emulating Warren's early page layouts and by publishing new work created in the classic style by original Warren artists, writers and editors.
Each issue of The Creeps is a highly collectible instant classic.
The Creeps 2019 Spooktacular recreates the iconic Warren horror annuals popular in the 1970's by emulating Warren's early page layouts. This 2018 Annual collects some of the best stories from the first two years of The Creeps in a perfect bound collector's edition. Each issue of The Creeps is a highly collectible instant classic!


text:
Being a bridesmaid: An honor that can get expensive
Suzanne Pollak
The since-kindergarten friend, the summer camp friend, the practically-sisters friend, the little sister, the sister-in-law, the college friend and the roommate will answer without hesitation. "Yes, I'm honored," they will say without fully grasping that the "bridesmaid" title comes with a Prada-sized price tag.
"It's so expensive and a huge commitment," Sara Wolf said, "Everything adds up."
As a student at University of Maryland with no steady income, being a bridesmaid is her second largest expense of the year, after tuition.
Wolf never realized how much it would cost to be a bridesmaid until she sat down with some old receipts after two of her best friends' weddings. For one wedding she had spent $150 on the dress, an extra $150 to build-up the dress modestly, $60 for the fabric, $130 on hair and make-up, $40 toward the hotel bill, plus shoes, bachelorette outings and a present for the couple.
"My mom tells me that for a wedding gift, especially if you're in the wedding, you should be spending at least a hundred dollars," Wolf said.
After doing the math, Wolf spent more than $700 for one wedding and combined $1,000 for two of her best friends' during the summer. Compared to the national average, Wolf's spending was on the lower side. Wedding and other websites like The Knot and Elite Daily reported that the average bridesmaid spends between $1,500 and $1,800 for the big day, including covering smaller events leading up to it, such as the engagement party, bridal showers and the bachelorette rendezvous.
College brides in particular, who have been bridesmaids themselves, understand the backstage costs; they often try cutting costs to minimize issues for their friends. For this reason, Avital Schwartz, a Kemp Mill native who wed in August at Congregation Shomrei Emunah in Baltimore, reserved the bridesmaid role for family.
"I hear how everyone else is talking about the stresses of being a bridesmaid and how it's so much drama," she said.
Schwartz feels that weddings are an important family milestone and giving friends such an involved job takes away from the family bonding experience.
As her family discussed wedding colors. Schwartz felt everyone should wear whichever color they liked best.
"I was totally flexible," she said.
Schwartz and her mother scoured eBay and found six pastel gowns with modest sleeves for $20 dollars each. The dresses needed $20 in hemming. Her bridesmaids did their own hair and make-up.
On the other hand, Wolf's experience dress-hunting to find a modest gown that matched the electric blue swatch she was given, was a "pain-in-the-neck."
"There were a lot of problems at the dress store because they don't sell extra fabric," she said. "Initially I thought I would buy a spool of fabric for a shawl, but the spool was a different color with mismatched stitching so I had to return it, which the [store clerks] weren't happy about."
In the end, Wolf purchased two dresses, one longer and one shorter so she could cut the shorter one for its identical material in order to tailor the longer dress.
"The shorter dress was originally $120, but they sold it to me for $60," Wolf said. "I told a friend and she tried to do the same thing, but they wouldn't give her the same deal."
Besides dresses, Schwartz's bridesmaids opted for smaller, low-key parties leading up to the big day to cut on costs.
"They made songs and poems and spoke at the parties and shower, she said. "I'm definitely happy in retrospect."
Newlywed Nini Slochowsky, who is pursuing a doctorate in clinical psychology at Long Island University, says she almost didn't have bridesmaids at her wedding because she was "scared of picking and choosing who was most important."
"A very wise friend of mine told me 'here's the secret point of bridesmaids. When someone gets married, you may or may not know the fiancé, you may or may not be living near them in the future, and all of a sudden you don't know where you fit into their new life. Here's a way for the bride to say, 'You're important to me and you'll always be important to me,'" Slochowsky said.
Ultimately, Slochowsky's sister convinced her to ask four people. Trying to be "minimalistic" and "intentional," the 24-year-old from Great Neck, N.Y., thought about the individuals she turns to most in her life.
Previous articleCake blends modern vegan trend with family heritage
Next articleNo Wagner for you
Planning an outdoor wedding? Learn from my experience.
Wedding vendors give back
Here are ways you can make the best of your wedding plans

redpajama_set_name: RedPajamaCommonCrawl


Anychart is a flexible Macromedia Flash based solution that allows you to create animated,compact, interactive and great looking charts. Driven by XML interface, it has No installation and Extremely Easy in Implementation in Web and in Applications. There is no need for Active-X on the server - graphics is rendered on client. All scripting languages (ASP, PHP, ColdFusion, Perl, etc.) are supported. Easily add your custom images, sounds, texts.


After your nightly rituals of washing your face and brushing your teeth, you head to bed. Healthy habits indeed. But do you also practice sleep hygiene? In this Mayo Clinic Radio Health Minute, Dr. Lois Krahn explains just what that means.


We're delighted to have collaborated with New Zealand's most trusted broadcaster, Mike McRoberts, to produce a new series of videos for ABC Business Sales.
Over the past couple of years, we've been working with ABC to develop a video-first content marketing strategy. The majority of business people are searching for opportunities on their mobile phones, so video is the best way to share a story effectively. And every business has a story.
Mike was involved in every element of the production process… here's the first episode in a series….


Introducing pyTower, a python based Sim Tower clone. This is a new project I started, after finding the existing clones lacking. So far it's in a really useless stage of development, but I wanted to show if what I had done so far.
I'm building it on the pygame library, which is a low level wrapper around SDL. I have to say, I am incredibly impressed with this library. You can get started really, really fast. Just focus on surfaces, rectangles, events and blitting. Ta-da, you have the basic tools for a game.
Anyway, the dev work is at github for your enjoyment.
One of the (I think) cool things I am doing is implementing the menu system in Qt4. Menu's are hard in pygame, IMHO, so I went around them. This gives it a bit of an MDI feel, but I like it so far. If things go well I might find a way to move the mini box into another window. I think this will be beneficial long-term because I'm planning on making the technology tree really flexible and extensible.
Anyway, here are the totally not-cool looking screen shots I have so far. Art is hard.
The main menu, Qt4 + pygame.
The game space, with mini map!


Arthritis wears away at your joints making every day activities extremely difficult. Nearly 27 million Americans suffer from osteoarthritis, which is the most common form of the disease. Dr. Geoffrey Westrich, an orthopedic surgeon at Hospital for Special Surgery in New York City, clears up the top misconceptions about the disease.


Can SwitchResX disable a display ?
SwitchResX includes a function to remove a display from the System's list of managed display. This means the display in question, once deactivated, will not be visible from the System or from any application. Only SwitchResX will be able to activate the display again.
Please also note that if you have mirroring enabled, the menu item will NOT show. So be also sure to disable video mirroring.
There are however some compatibility problems on Yosemite and Mavericks: see this section of the FAQ.


With this motto in mind, we've got become certainly one of by far the most technologically innovative, cost-efficient, and price-competitive manufacturers for Asphalt Cutting Blade , asphalt cutting blade , Diamond Asphalt Cutting Blade , providing customers with personalized service and unique products.
"Sticking to your principle of ""Super Good quality, Satisfactory service"" ,We are striving to get a fantastic business enterprise partner of you for Asphalt Cutting Blade , asphalt cutting blade , Diamond Asphalt Cutting Blade , We've got constructed strong and long co-operation relationship with an enormous quantity of companies within this business in Kenya and overseas. Immediate and professional after-sale service supplied by our consultant group has happy our buyers. Thorough Info and parameters from the merchandise will probably be sent for you for any thorough acknowledge. Free samples may be delivered and company check out to our corporation. n Kenya for negotiation is constantly welcome. Hope to get inquiries type you and construct a long-term co-operation partnership.


What to get involved? Looking for ways to BE the church in the world. Take a look at our ministry opportunies.
We believe that we are on thie journey together. No matter who you are or where you are on life's journey, you are welcome here. Our Community Formation ministry is about living together in community where we provide support and prayer for each other and the world.
Our Evangalism and Hospitality ministry exists to carry God's extravagant welcome to a hurting world. If you have a heart to feed the hungry, work to end oppression, and be the love of God in our world then the Evangalism and Hospitality ministry might be for you.


Made in the USA, the Multi-Function Leash by Blue-9 can do it all. With 3 high-strength D-rings and 2 snap hooks, this Jack-of-all-trades can be used as a 6 foot, 4.5 foot, 3.5 foot and hands-free leash. Additionally, when paired with your Blue-9 Buckle Neck Balance Harness, attach both ends to the front and rear D-rings for dual control of your pet. Whether you want to free up your hands, give extra distance, or find yourself needing to shorten things up in a busy situation, this leash provides you with options.


Update your summer wears with the Joules Cruise long chino shorts.
Crafted from a cotton rich blend with a touch of stretch for the ultimate fit and comfort. Cut to a classic style with front and back pockets and are cut to a slightly longer length for a relaxed vibe.
Tuck a white tee into these and wear with pumps ready for a weekend full of fun and adventure.


LEXINGTON, Ky. – The University of Kentucky women's basketball team's full 2018-19 schedule is set as the Southeastern Conference has announced the Wildcats' 16-game league slate, which includes home games against Vanderbilt, Ole Miss, Missouri, South Carolina, Florida, Arkansas, LSU and Texas A&M.
On top of its eight home games, Kentucky will travel in the league to play at Mississippi State, Tennessee, LSU, Texas A&M, Auburn, Alabama, South Carolina and Georgia. The SEC schedule is a challenging 16-game slate, playing every team in the league at least once and three teams twice in one permanent opponent (South Carolina) and two opponents (LSU and Texas A&M) that rotate annually.
For the second year in a row, Kentucky will begin its conference slate at home, squaring off with Vanderbilt on Jan. 3 in Memorial Coliseum before back-to-back road games at Mississippi State on Jan. 6 and Tennessee on Jan. 10. After returning to Memorial Coliseum to face Ole Miss on Jan. 13, UK travels for its first of two games during the year against LSU on Jan. 17 before a bye date on Sunday, Jan. 20. The Wildcats will return from the bye with a home game vs. Missouri on Jan 24 before a road showdown at Texas A&M on Jan. 27. UK concludes January by hosting South Carolina inside Memorial Coliseum on Jan. 31.
February starts with the Wildcats hosting Florida on Feb. 3 inside Rupp Arena before a two-game road stretch at Auburn on Feb. 7 and at Alabama on Feb. 11. Following another bye date on Feb. 14, Kentucky will conclude the SEC slate with three of its last five at home starting with a game vs. Arkansas on Feb. 17. After traveling to South Carolina on Feb. 21, UK will host its annual Senior Day game on Feb. 24 vs. LSU before its final home game of the year on Feb. 28 vs. Texas A&M. UK concludes its regular season on March 3 at Georgia.
Game times for all 11 of Kentucky's non-conference home games, including its lone exhibition of the year, have been set, while the UK-Morehead State game originally scheduled for Thursday, Nov. 29 has been moved to Wednesday, Nov. 28 at Noon ET. In addition, Kentucky will play non-conference foe Virginia inside Rupp Arena on Thursday, Nov. 15 at 7 p.m. ET. Complete game times are listed below.
Season ticket renewal invoices are available online at UKathletics.com/renew. Each season, season ticket holders are able to request a seat upgrade upon renewing their season tickets. Seat upgrades are based upon availability. To request an upgrade, fill out the upgrade request section through your online renewal invoice or write the request on your hard-copy season ticket renewal invoice, which was mailed out in early August. Upgrade requests will be reviewed after the season ticket renewal deadline.
Earlier this summer, UK Athletics announced that a new, state-of-the-art video board will be installed at the south end of Memorial Coliseum. The new video board will be located above the front doors at the Avenue of Champions entrance. A full-feature ribbon board will be installed at the north end of Memorial Coliseum, just below the balcony, and the existing north end video board will be removed. These upgrades are the first steps in plans to enhance the fan experience in Memorial Coliseum.
During the week of January 20-27, SEC member institutions will participate in the seventh annual "We Back Pat" Week. The week-long initiative is focused on bringing awareness and recognition to the Pat Summitt Foundation, a fund of East Tennessee Foundation, and its fight against Alzheimer's disease.


Sydney performs with Octavia, a professional eight-harp ensemble which produced the CDs "Angels We Have Heard On High" and "Moonlight Sonata". She played with the Fort Worth Symphony for many years, and is a guest performer with many other instrumental ensembles. She performs at many venues throughout the Dallas-Fort Worth area.
Sydney specializes in weddings and plays many solo performances. Often brides will want to listen to suggested musical selections in her home studio in West Ft. Worth TX. She plays traditional wedding music as well as classical, love songs, show tunes, and Christian and Hebrew songs. She is available for weddings, funerals, receptions or special programs, including lecture demonstrations of the harp. To schedule a performance or for more information, contact Sydney.


The newest addition to Vitus' city bike lineup, the Vee 29 is a smooth-rolling big-wheel commuter with reliable components and finishing kit.
An aluminium city bike with 29er wheels mean a bike that rolls better over rough roads and small kerbs. Complete with Promax V-brakes and a single speed drivetrain, the Vee 29 City Bike is the complete cruiser for the style conscious and the practical commuter.


A quick content about Beautiful watercolor tattoos . Read more.
Thoughts on Beautiful watercolor tattoos are below.
Start with thinking about why you'd like the tattoo. There are a lot of choices and inspirationA to select from the original tattoo tips for men that I've collected for you. The right sleeve tattoo ideas can be extremely elegant, whereas the incorrect sleeve design may resemble an enormous mess.
Tags are Beautiful watercolor tattoos , tattoo, marks.


Dedicated to improving the homes of military families all over the U.S., the award-winning show, Military Makeover, invites CertaPet to join the cast and crew of its 17th upcoming season.
Alongside host, Montel Williams, we got the opportunity to sit down, talk, and work with a special family whose lives were changed by an emotional support animal. As a veteran-owned company, CertaPet understands the time and hardship that goes into the lives of servicemen and women as well as their family so we are grateful we get to be a part of this experience.
"We are so honored and excited to be working with Military Makeover and providing our services to military families. As a military spouse working for a veteran-owned company, it was a very natural alignment." said Prairie Conlon, CertaPet Clinical Manager.
On this new season, the featured Military Makeover Family will be the Middletons. Beyond being sponsorship, CertaPet assisted them with clinical assessment and consultation with their Emotional Support Animal.
With the help of CertaPet and other well-known brands, Military Makeover will offer hope and a helping hand here on the home front to members of this family.
"Military families are consistently on the move. This means changes in work, schools, environment, family life, friends and social life, etc. The family pet is a consistency that brings a sense of "normal" to the multiple homes throughout all these changes." Prairie states.
Military Makeover episode 3 airs March 15, 2019. We make a special appearance!
Emotional support animals are more than just pets. They're a treatment to your mental health and ultimately, your furever family.


At DumpsterFly, we strive to bring you the best service at the best price available. We have a wide selection of dumpster sizes in stock in Marrowbone, KY, with flexible & timley pickup and delivery. Give us a call today for all your dumpster rental and roll off needs.
Another reason your parents may be moving out of their home is they want to move to a different state. Possibly in your mother's or father's golden many years someplace warmer intrigues them. Or perhaps they want to turn to the country to enjoy more peace and quiet. The metropolis may fascinate your parents since they will not have drive an automobile as much and they can get his or her exercise when walking to the supermarket or dining places that will revolve around them.
With regards to a car port that needs clearing out, chances are excellent that there is a pretty huge amount involving junk being thrown away. Since a lot of community trash organizations impose limitations on how very much trash place out weekly, and a lot of points may just be too big and unclearly shaped for trash hand bags anyway, you're going to want to rent the dumpster from a reliable Rent a Waste Management Dumpster in Marrowbone, KY company. Just picturing the dumpster-sized load involving junk absent from your car port should make the task at hand seem far more manageable.
In hearing the phrase prevention of contamination your mind conjure up while using idea of mitigating this emission involving fossil fuels produced by the travelling of automobiles. It's a good indicator, but let me tell you, the term Smog is not aimed at the air contamination and sound pollution that's generated from automobiles. There are ton many other variables contributing to contamination of the natural environment in the cities such as development waste, natural yard throw away, industrial throw away.
Check dumpsters in apartment complexes and the starting point and conclude of thirty day period. When new tenants proceed in/out they outdoor tents to throw away many things of worth.
Most people take a look at renting a dumpster just as one expense. This particular cannot be turned down, as it is expensive to borrow the dumpster from a corporation. However, men and women rarely evaluate the savings in which using a dumpster can supply. Businesses typically use them, simply because dumpsters are the well organized way to dispose of a large volume of garbage. Nevertheless, a dumpster may ease the worries on an person's pocketbook. Listed below are three ways dumpsters help save people dollars anytime they will take on huge projects.
It is just a good indicator that you have taken a decision to manage waste created in an Eco- helpful manner nevertheless, important undertaking here is the technique you plan to be able to dispose this generated throw away. Trash created in the projects like development, house redevelopment, industrial projects and natural yard cleaning is very big. Planning to get rid of the throw away on self applied makes you tumble sick with a few injuries when not properly managed. Handling this kind of huge a great deal of waste requires expertise and professionalism. That even will kill your time simply because all the garbage will be managed manually.
Eventually, once you are finished using the rented dumpster, a call for the rental company for the pick-up is required. Many companies have got different rental intervals, and having the number of days allowed for a rental is definitely imperative to understand before making a repayment.
Once you've got your house decluttered and organized, you should keep a control out to your bad habits. Your own cluttered chaos didn't happen overnight once, and it will little by little creep back in to your lifestyle if you aren't very careful! Throw away pre-approved offers when it comes throughout. Keep your household tidy and organized. Ask yourself from time to time if you actually need some of the things that have been gathering dust. With a little extreme caution, your home will always be clutter-free.
Once the rummage purchase is over, you continue to may have a lot of junk to get rid of. In the past you thought you would use these items nevertheless it turns out you don't ever did. Certainly before the search sale or maybe Rent a Waste Management Dumpster in Marrowbone make sure there is nothing in your barn or maybe outer structures that your young children want. Allowed them to look over points and give these to them to retain. Now the things you do not use and want are generally out of your barn as well as garage.
Do you wish ones nation to face among the best nations on the planet? If yes, subsequently motivate yourself in promoting recycling in the waste in Eco-friendly manner at this time there by reducing the outcome of pollution on the environment. Not everybody which generates the waste is aware of the process of recycling, so the customer is in issue once again at this juncture. Expert Rent a Waste Management Dumpster in Marrowbone service providers in the market offers spellbound companies making your task hassle free with very affordable cost. These qualified Rent a Waste Management Dumpster in Marrowbone service providers just as one expert in the field has a due wish to ensure that the disposed waste can be recycled in environmental pleasant manner. Carrier's networks provide dumpsters on rental basis to be able to ward off ones generated squander and handle the post accountability of recycling the squander Eco-friendly manner.
One of these is to determine the company can provide you companies within the quickest time achievable. If you are developing or upgrading a creating, you would wish everything to advance fast plus in a synchronized manner. If you are constructing a household that is created for business, you would want it to become finished in time so that clients move in. It is for this reason that you need a new Rent a Waste Management Dumpster in Marrowbone company that will always be about to give companies and get any project moving. Consequently, consider this because the more time you waste, the greater money you may well lose.
Once you have a big clean-up job ahead of you, renting a dumpster from a Rent a Waste Management Dumpster in Marrowbone company could make the job much easier. However, as with every company you hire that may be coming to your home, you need to be able to trust them. Here are a couple issues that you may be concered about, and why going with a small, local company would be the solution to these people: Ideal for Home enthusiasts or long-term projects, Rent a Waste Management Dumpster in Marrowbone can make it an easy task to collect your junk in one location for easy crap hauling. Pertaining to do-it-yourselfers who don't intellect a little work, Rent a Waste Management Dumpster in Marrowbones eliminate the hassle as well as added time and financial valuation on hauling weight after weight of rubbish for the dump with your sedan. Rather than making vacation after vacation, you can just drop all of your undesired items--including bulky household furniture or home appliances that probably would not fit in ones car--right into the dumpster. When you find yourself done, the dumpster company may remove the container--and ones junk--in one easy stage. And because quite a few offer storage containers in various sizes, ones rental can be custom-made for your unique project, assisting keep your cleaning as cost-effective as is possible. Roll-off Rent a Waste Management Dumpster in Marrowbone also helps corral squander in one place, rendering it great for having on-site during the amount of a remodelling or long-term house cleanout.
One of the primary benefits of dumpsters is you can fill these with whatever kind of trash you might have. Think about that before you even check into Rent a Waste Management Dumpster in Marrowbone, KY prices. No matter if you need to utilize them for appliance removal, furniture removal, or just simple junk removal, if you can fit it in that room, you can put the item in there. It is great news if you're doing a overall cleanout of a property, garage, or another building. Traditional garbage removal possibilities don't present you with this luxury--you get to take your extra-large items to a transfer place or dump on your own. Which has a dumpster rented from your local crap hauling company, though, you are able to dispose of nearly anything you want to.
It is no lengthier difficult for an individual to fetch the garbage cans the best suit your needs. Dumpster rentals are generally priced depending on the tonnage of waste, period of rental and also size of dumpsters You'll be over weighted down by further charges with regards to multiple trips to dispose your own trash when you hire a small size dumpster. You're paying for the disposable space when you hire a significant size dumpster. Most of the Rent a Waste Management Dumpster in Marrowbone, KY companies out there put a finish to this problem if you rent then different sizes regarding dumpster according to your own customized needs.
Renting a dumpster can be a lifesaver when you have a significant project on-going at home. These kind of rentals are great for trash removing, furniture removal, and crap hauling, among other things. However, it is vital to remember that hiring a dumpster does not automatically imply your dumpster master will be in the grass to move the garbage into the container for you. Ensure you know exactly what's included ahead of time when you rent a dumpster so that you really don't run into virtually any surprises on the way.
Good, you have chosen to rent a dumpster to clear the a lot of trash accumulated at your work location. You should consider the type and also size of dumpster that best fits your requirements before you go in advance with the rental course of action. Roll away from dumpster should be your choice if you are looking out to dump big tons of waste in the main commercial and residential assignments. Before you place a purchase order for the move off waste bin, it is crucial for you to are aware of the dimensions of this garbage container so that you can accommodate the required place to squeeze dumpster at your undertaking location. You may be running a home renovation, commercial undertaking or manufacturing clean up, the place you support for the garbage jar should be double size of the height and also width.
In the beginning you may want to consider to have a rummage sales. Make the rates on the goods very low so you can just eliminate. Do not try to earn money at your rummage sales, you are obtaining it to get rid of things. It is advisable to sell your own items for affordable to someone that may use them or fix all of them than to only throw all of them in the dumpster and also end up in a junkyard.


text:
Utkání hvězd české extraligy, později Utkání hvězd extraligy ledního hokeje byly exhibiční zápasy ledního hokeje, hrané od roku 1996 do roku 2006 v české později i slovenské extralize. V roce 1996 neodstartovala pouze zlatá éra českého hokeje, ale také nová tradice, jež nese název Utkání Hvězd Extraligy ledního hokeje. Každoročně si nejlepší hráči působící v české nejvyšší soutěži (od roku 2003 i hráči působící v slovenské nejvyšší soutěži) dali o jednom z víkendů volno, aby sehráli divácky velmi atraktivní klání, které je proloženo, podobně jako v zámoří, soutěžemi zručnosti.

Na sestavování týmů se aktivně podíleli všichni, kteří hokej milují. První formace volí fanoušci, druhé novináři, třetí a čtvrté pak určí samotní trenéři.

Utkání hvězd mělo své příznivce i odpůrce: těm se nezdálo až příliš vlažné tempo exhibičního utkání i to, že hokejisté nešli naplno do dovednostních soutěží! Nicméně utkání hvězd bylo součástí hokejového kalendáře, odehrálo se desetkrát (do roku 2006).

Jednotlivé ročníky

Rekordy individuálních soutěží

Česká hokejová extraliga
Lední hokej v Česku
Sportovní události založené roku 1996

redpajama_set_name: RedPajamaWikipedia


text:
Boeing's Chadwick: F/A-18 line could run beyond 2017, Bloomberg reports
November 18, 2020 October 31, 2014 by iam387
Boeing's defense chief remains optimistic about the F/A-18 fighter assembly line in north St. Louis County, saying there's a good chance the line could stay open beyond 2017, according to Bloomberg News.
Current orders will keep the assembly line open until 2016. If approved, a Congressional proposal to buy 12 EA-18G Growlers, an electronic warfare version of the Super Hornet carrier-based fighter, would allow Boeing to keep the line up running end of 2017.
However, the aircraft maker has more than a "50-50 chance" of receiving enough Navy and foreign orders to keep the Super Hornet line open beyond that date, Chris Chadwick, president of St. Louis County-based Boeing Defense, Space and Security, told Bloomberg News today.
Guide Dogs Raffle


How To Apply: To apply Visit www.worksafebc.com and select Careers to submit a resume and cover letter that details your characteristics, accomplishments, and prior work experience related to the requirements for the position. We can only accept applications submitted through our website.
We have an opportunity for a family and critical response specialist to work with a dedicated team that provides support to workers, family members, and employers who have experienced a traumatic workplace accident, and Prevention and Investigations officers who respond to these incidents. Consider joining our team in Richmond for this one-year assignment to make a difference in the lives of workers and employers across the province.
At WorkSafeBC, we're dedicated to promoting safe and healthy workplaces across B.C. We partner with workers and employers to save lives and prevent injury, disease, and disability. When work-related injuries or diseases occur, we provide compensation, and support injured workers in their recovery, rehabilitation, and safe return to work. We also work diligently to sustain our workers compensation system for today and future generations. We're honoured to serve the 2.3 million workers and 231,000 registered employers in our province.
Visit www.worksafebc.com and select Careers to submit a resume and cover letter that details your characteristics, accomplishments, and prior work experience related to the requirements for the position. We can only accept applications submitted through our website.
Thank you, in advance, for applying. Unfortunately, we're only able to contact those whose applications we will be pursuing further.


Arrummula Constructions is a known name in the Real Estate market. With years of experience in the construction and development field, the Group is known to be the best Real Estate developer in the city. The builders believe in providing high quality construction work to all home seekers helping them achieve their dream of owning their own home. To preserve a strong grip in the highly competitive real estate market, its extremely trained team focuses on offering excellent realty service to all customers. One of the top builders in the city, Arrummula Constructions strongly believes in giving the best infrastructure to its clients without compromising on the quality to offer a fine living space.
Start a new discussion about Arrummula Constructions!


Elders and teachers often teach us to do work in symmetry. There is actually a Mathematical segment that sheds light on this aspect. We know it as Geometry. Commonly, it is the classic story of lines, curves and angles but there are deeper shades to it.
In fact, the permutations get so complicated at length that if you are not too indulgent or passionate about the segment, you may find it overbearing. In this case, you will require external assistance in sorting out the homework.
Parents and neighbors – You can approach them in your blue days and ask for help regarding Geometry. They may be well-heeled with the segment and may help you exponentially. It helps if you have a retired Math teacher living in your locality.
Classmates – You can ask classmates as to how to solve particular questions and where to seek answers for the same. They can guide you with pertinent sites or reference books that are quite efficacious. Some students actually take active interest in Math and Geometry.
Math sites – You can place your questions on the Math sites and the answers may come floating in. Quite a few of these sites start off with free services. When they become famous, they obviously begin to charge for the same. Keep an eye.
Crisp videos – You can check out pertinent videos on video sharing sites. Some are started by worthy Geometry teachers, sharing their insights and giving effective tips on tackling the subject. You need to pay attention to the details. The rest follows.
Social media site – This is where you get many learned people willing to help you. You can also try educational forums and get enlightened. Some even get thrilled when asked to solve questions. The subject actually interests a few substantially.
DVDs and Podcasts – You may also use the stratagem of seeing relevant DVDs or listening to Podcasts on the subject in your free time or at the time of sleeping. You will come across brilliant ideas an innovations, id the products are smartly cut. In all fairness, however, this homework solution is not free as you need to buy the DVDs and Podcasts.
The above-mentioned pointers should help you in your quest.


The role of transposable elements in sculpting the genome is well appreciated but remains poorly understood. Some organisms, such as humans, do not have active transposons; however, transposable elements were presumably active in their ancestral genomes. Of specific interest is whether the DNA surrounding the sites of transposon excision become recombinogenic, thus bringing about homologous recombination. Previous studies in maize and Drosophila have provided conflicting evidence on whether transposon excision is correlated with homologous recombination. Here we take advantage of an atypical Dissociation (Ds) element, a maize transposon that can be mobilized by the Ac transposase gene in Arabidopsis thaliana, to address questions on the mechanism of Ds excision. This atypical Ds element contains an adjacent 598 base pairs (bp) inverted repeat; the element was allowed to excise by the introduction of an unlinked Ac transposase source through mating. Footprints at the excision site suggest a micro-homology mediated non-homologous end joining reminiscent of V(D)J recombination involving the formation of intra-helix 3′ to 5′ trans-esterification as an intermediate, a mechanism consistent with previous observations in maize, Antirrhinum and in certain insects. The proposed mechanism suggests that the broken chromosome at the excision site should not allow recombinational interaction with the homologous chromosome, and that the linked inverted repeat should also be mobilizable. To test the first prediction, we measured recombination of flanking chromosomal arms selected for the excision of Ds. In congruence with the model, Ds excision did not influence crossover recombination. Furthermore, evidence for correlated movement of the adjacent inverted repeat sequence is presented; its origin and movement suggest a novel mechanism for the evolution of repeated elements. Taken together these results suggest that the movement of transposable elements themselves may not directly influence linkage. Possibility remains, however, for novel repeated DNA sequences produced as a consequence of transposon movement to influence crossover in subsequent generations.
Copyright: © 2007 Langer et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: Work in AR's laboratory was supported by National Science Foundation grants IBN-9982414, EIA-0130059, and EIA-0205061 to AR. Work in UG's laboratory was supported by the Cold Spring Harbor Laboratory President's Council and the European Molecular Biology Organization grants to UG.
Mobile genetic elements are important forces of evolution . Ac/Ds elements are DNA transposons that use a conservative "cut-and-paste" mechanism involving double-stranded DNA breaks to move within the genome. Repair of broken chromosome ends at the donor (excision) site contributes to genetic alterations in adjoining sequences –. Footprints or DNA sequence rearrangements left at the donor site following transposition provide clues to the repair mechanisms , , .
The exact mechanism of donor site repair remains unresolved; indeed, it appears likely that this phenomenon may be affected by host proteins and hence might vary among species . In particular the effect of Ac/Ds transposition on chromosome crossover, a major mechanism behind genetic diversity , is an unanswered question. Linkage disequilibrium, the historical evidence of rare crossover exchanges between close markers, may potentially have its origin, among other factors, in rare crossovers stimulated by the activity of mobile elements. Early studies in maize of Activator/Dissociation (Ac/Ds) elements had failed to find a clear association between crossing over and transposition , . Although transposons are frequently involved in gene conversion  and stimulate crossover within repeated DNA , recent genome level analysis shows either no correlation  or negative correlation ,  between recombination rate and transposable element density along the chromosome. One may speculate that this may be the consequence of evolutionary selection to prevent crossover exchanges where transposon density is high. The question remains whether a pioneering transposable element in a chromosome free of prior copies of that transposon allows high frequency crossover during its movement.
To address these questions, here we examined transposition of an active Ds element artificially introduced into Arabidopsis thaliana. The Ds element has an atypical insertion site, in which there is a long unrelated inverted repeat DNA sequence located immediately adjacent to, but outside of, one of the terminal inverted repeats, which is bordered on the outside by one of the duplicated target sequences. As in previous reports , ,  we show that the excision repair footprints support a mechanism resembling immunoglobulin V(D)J class switching in vertebrates , . In accordance with the proposed model of Ds transposition, we show that transposition does not influence crossover recombination between homologous chromosomes. This result suggests that the movement of transposable elements may not be an important factor in genome rearrangement by recombination. Furthermore, as predicted by the model, we detected movement of the unrelated DNA inverted repeat from its initial position to secondary locations. These observations, along with an analysis of the footprints of transposition, provoke a new model of DNA repair at the excision site that allows the formation of new inverted repeat sequences adjacent to the transposable element. We speculate that such inverted repeats may subsequently separate from the original transposon and form novel mobile sequences.
The Ds site from which excision was allowed has an atypical configuration at the 3′ end: the 3′ terminal inverted repeat (3′ TIR) of the Ds end is situated adjacent to another inverted repeat of 598 bp (Figure 1A). The sequence of this repeat was derived from the parental strain  in which the maize Ds element, introduced into Arabidopsis on a T-DNA vector, had previously transposed into the current donor site. The presence of the 598 bp repeat suggests that this duplicated sequence transposed into this site from its original location on a T-DNA stably incorporated into the chromosome in its parent. (Figure 1B). Although the 5′ end of the Ds insertion has the typical half-site of a target duplication, the other target half-site is separated from the Ds TIR by the 598 bp inverted repeat sequence derived from the T-DNA vector. To test whether this modified Ds element can transpose, we mobilized the element by introducing an active Ac transposase gene as described below.
Figure 1. Structure and position of the Ds element in strain ET500.
(A) Sequence of the Ds element and adjacent sequences showing the important elements. The insert is flanked by 7 bp target site duplication (boxed). DNA outside the duplicated target sequences are identical to the surrounding plant DNA sequence. Sequence of the 598 bp inverted repeat is identical to a fragment of the T-DNA containing 3′-octopine synthase promoter (3′OCS) and the right border (RB) element. Black circle within the boxed sequence shows the site of sequence inversion. (B) Structure of the Ds element. The Ds element contains a promoter-less bacterial GUS gene just inside the leftward TIR, and a bacterial NPTII gene with its own promoter. A plant-specific promoter-enhancer element (stippled oval) 5′ to the Ds insertion site confers constitutive GUS expression. (C) Map of the chromosomal region containing Ds. Numbers denote the first base on chromosome 4 (counting from the top) for the respective site. Roman numerals denote the two genetic intervals assayed for crossover. The transposase source, Ac, was unlinked to chromosome 4, and was present only in the experimental cross. Superscripts identify La-er (L) and No-0 (N) alleles.
We crossed two strains of Arabidopsis thaliana, Landsberg erecta (La-er) and Nossen (No-0) such that the F1 provided heterozygosities at molecular markers for scoring recombinants (Figure 1C). The experimental cross brought an enhancer detector Ds element from one parent together with an unlinked and immobile Ac transposase gene from the other. The Ds contains a beta-glucuronidase (GUS) reporter gene and a neomycin phosphotransferase (NPTII) marker located within bounds of the two TIRs (Figure 1B), while Ac is adjacent to the negatively-selectable TMS2 gene encoding an amide hydrolase that imparts sensitivity to naphthalene acetamide . The F1 of the corresponding control cross in the same genetic background carried the same Ds but not the Ac. The GUS reporter gene was under the control of an endogenous enhancer leading to constitutive expression such that production of GUS− somatic sectors could serve as the hallmark of Ds excision. The control cross did not show any evidence of somatic Ds excision (32 F1 plants tested) as indicated by a lack of GUS− leaf sectors (Figure 2A), nor of germinal excision as evidenced by transmission to the F2. By contrast, high frequency Ds excision occurred in 47 of 53 F1 plants of the experimental cross, as evidenced by GUS− sectors on F1 leaves (Figure 2B) and confirmed by polymerase chain reaction (PCR) and Southern analysis following transmission into the F2 progeny (Figure 2C). To prevent continued transposition during analysis, F2 plants were selected on naphthaline acetamide (NAM) to eliminate descendents inheriting Ac.
Figure 2. Excision of Ds.
(A) A leaf of a parental Ds strain homozygous for Ds (ET500), showing uniform GUS+ staining. (B) GUS− somatic sectors due to Ds excision in F1 of ET500×NaeAc. (C) Southern blot analysis showing the excision of Ds. Upper panel was probed with GUS; lower panel represents the same blot cut off between 2kb and 3kb markers, and probed with DCL1 cDNA sequences as a loading control . Lane 1, ET500 parental (before Ds excision). Lanes 2-3, DNA from two independent F2 derivatives of ET500×NaeAc showing Ds excision, and lane 4, DNA from an F2 plant showing excision and reinsertion of Ds to another location. (D) Amplification of molecular markers: Landsberg erecta (L); Nossen (N); heterozygous (L/N) Left hand panel, nga8; middle panel, Ga1.1/BsaB1; right hand panel, nga1111.
Depending on the timing and relative rates of Ds excision in the F1, meristem cells contributing to the germ line may contain numerous independent transposition events or one early event with numerous clonal descendents. Thus, a single F2 family from one flower could represent multiple independent excision events or one clonal event. A sampling of independent events could not be guaranteed by analyzing a number of F2's derived from a single F1 plant. To overcome this difficulty, from each F1 parent only one F2 progeny plant was scored for recombination. Each F2 plant was selected for analysis at random from among a pool of F2 plants that was proven to have an excision event by PCR analysis of the donor site and confirmed by Southern blots. Each separate pool was derived from a single F1 parent. This guaranteed that each F2 plant examined represented an independent Ds excision. Although it was not possible to formally distinguish between premeiotic, meiotic or postmeiotic transposition, most transposition probably occurred premeiotically because of the observed high rate of excision plants per F1 line. There was no attempt to determine whether crossover events were somatic or meiotic.
DNA sequences at 11 of 17 independently derived empty Ds donor sites showed the same footprint and at three others a single base difference from the commonest footprint (Table 1). Sequences remaining at the excision sites were most consistent with the idea that excision frequently involved only one of the duplicated target sequences of the original Ds integration site. The footprints suggest a micro-homology mediated non-homologous end joining mechanism reminiscent of V(D)J recombination as suggested for Tam3 excision in Antirrhinum , Ds excision in maize ,  and Arabidopsis , and shown for in vitro transposition of the insect hAT element Hermes  and in vivo excision of Ds in yeast , , involving pairs of staggered nicks, followed by DNA synthesis, hairpin formation by trans-esterification, hairpin nicking, imperfect base pairing, ligation, and mismatch correction (Figure 3A). Essentially similar models can explain two additional footprints (Figure 3B, C), and the remaining one involved a 41 base pairs deletion (Figure 3D). The exact site where the nick distal to the Ds element occurred cannot be ascertained from the footprint result; however, previous studies in maize, Arabidopsis and tobacco , , , , ,  have determined that the footprints are most consistent with the transposase-mediated cleavage occurring at the 5′ nucleotide flanking the insertion. Our results are consistent with the previous models for 16 of 17 footprints (Figure 3). For the remaining footprint, involving a 41 bp deletion (Fig 4D), a mechanism involving an exonuclease-mediated degradation starting at a position 5′ of the Ds element is plausible, consistent with studies that show exonucleolytic deletion of sequences flanking the end of an element , . However, the model we have proposed here (Fig. 3) explains all footprints observed here, including footprints reminiscent of V(D)J recombination, a footprint involving the 41 bp deletion and the formation of a 598 bp inverted repeat adjacent to the 3′ TIR of the starting Ds used in this study.
Figure 3. Models explaining Ds excision footprints.
(A–C) A model explaining the formation of the most common footprints. Boxed bases are the 7bp duplicated target sequence. Thin straight arrows show putative transposase nick sites, curved arrows depict trans-esterification reaction between the 3′ OH group of one strand and the 5′ phosphate of the other, and arrowheads show the points of ligation. Gap repair synthesis bases are in bold and direction of synthesis is shown by an arrow; deleted bases are underlined. Note the absence of symmetrical targets flanking the excision site. (D) Model explaining the formation of a less common footprint.
The model suggests two independent predictions. First, the proposed mechanism of restoration of the broken chromosome does not involve any opportunity for homologous recombination initiated by the DNA double strand breaks because the broken ends are trans-esterified into hairpins, therefore, cannot initiate double strand break repair  without further processing. The second more subtle prediction concerns the observed 41 bp 'sliding' of the transposase to produce the second nick (Figure 3D). It is conceivable that a slide by the transposase could make a double chain break at a site distant from the 3′TIR, upon which the new end could undergo hairpin formation, followed by a second nick adjacent to the 3′TIR (Figure 4). As a result, a new inverted repeat sequence could form, as explained in Fig. 4. While this prediction is actually prompted by the observation of the 3′ inverted repeat at the original Ds element site (Figure 1A), it also suggests that a reversal of the process may occasionally co-mobilize the inverted repeat sequence along with the Ds element. Tests of these predictions are as follows.
Figure 4. A model proposing the formation and movement of novel inverted repeat elements.
(A) TIRs of a transposable element are shown in filled boxes. The boxed sequences are the 7bp duplicated half-sites. Transposase molecules bind at the TIRs, and (B) make pairs of staggered nicks on each flank (triangles). Occasionally, the DNA may slip (C) or the transposase may bind to cryptic sites on adjacent host DNA, and make a paired nick, which leads to excision (D). (E) Short repair synthesis followed by trans-esterification leads to a hairpin at the host segment attached to the TIR, which is nicked again near the TIR. Helix unwinding, followed by microhomology-mediated ligation to a new host site inserts the transposon, and single strand gap filling by repair synthesis forms a novel inverted repeat derived from host sequences adjacent to the original location (F). The transposon may repeat the cycle of precise (B) or imprecise (C) excision, and may be dissociated from novel host-derived inverted repeat (G). If the host-derived sequence had a cryptic transposase-binding site, the site is now duplicated on the novel inverted repeat. This may lead to independent movement of the inverted repeat sequence as a novel mobile element so long as a transposase is available.
Among F2 seedlings from the experimental cross Ac (No-0)×Ds (La-er), we selected progeny that had experienced a Ds excision in the following way. We required that each F2 plant lacked Ac, was uniformly GUS−, and had at least one La-er allele of nga8 (nga8L) tightly linked to Ds (Figure 1C). These plants contained at least one chromosome from which Ds had excised from the donor site. Single plants representing independent events from each of 47 F1 families from the experimental cross were identified, and analysed by PCR for crossover within Ga1.1—nga8—nga1111 intervals (Figure 1C). Of 47 independent excision lines, 8 had crossover recombination between nga1111 and Ga1.1 (Table 2). This recombination rate (8.5% per chromosome) is not significantly different from 10% recombination per chromosome in the control cross where no transposition was possible (29 crossovers among 144 F2 plants tested). Given 10% spontaneous crossover rate, 17 or more crossovers per 47 Ds excision events would have been scored as significantly correlated (χ2 = 4.98; P ≤ 0.05, for 1 degree of freedom). Thus, the ability to detect an increase in crossover rate to 18% or higher argues that we would have detected a 50% crossover rate predicted by the double strand break-repair mechanism  had the repair following Ds excision occurred by the strict version of this mechanism. Crossover frequency below 18% is presumably due to events unrelated to Ds excision.
A critical condition of the above test was that selection on NAM did not influence segregation of the three markers flanking Ds, namely, nga8, Ga1.1 and nga1111. We tested this by crossing the Ac (No-0) parent to wild type La-er, and analyzing marker segregation in F2 lines that were either selected or not selected on NAM. For 26 F2 plants not selected on NAM, the number of plants segregating homozygous Nossen alleles (NN): heterozygous (NL): homozygous Landsberg alleles (LL) were 9∶12∶5 for nga8, 6∶15∶5 for Ga1.1, and 8∶13∶5 for nga1111, respectively (see Figure 2D for a typical result). For those selected on NAM, the corresponding numbers were 9∶17∶1 for nga8, 11∶13∶3 for Ga1.1 and 8∶17∶2 for nga1111. The pairs of segregation ratios of the selected versus unselected plants were indistinguishable from each other by the chi-squared test (P≤0.2 for nga8; P≤1.0 for both Ga1.1 and nga111, respectively). In these crosses, crossover frequencies in Ga1.1—nga8 and nga8—nga1111 intervals were 3/52 chromosomes and 1/52 chromosomes, respectively, in the presence of NAM selection, and 4/54 chromosomes and 2/54 chromosomes, respectively, in the absence of NAM selection. These pairs of frequencies in selected versus unselected lines are statistically indistinguishable. Moreover, for 231 F2 progeny of 53 families screened for Ds excision (selection on NAM for Ac−, GUS− phenotype) the ratio of LL:NL:NN was 24%∶45%∶31%, showing no significant deficiency of any class with selection on NAM. The TMS2 locus is unlinked to nga8, satisfying another requirement of the experiment. Linkage of TMS2 (N) to nga8N would have resulted in a deficiency of NN at nga8 with negative selection on NAM. PCR confirmed that NAMS plants were also Ac−.
The regions flanking the excision footprints (Figure 5 A, B) were examined by Southern analysis to determine the fate of the 598bp repeat after Ds excision (Figure 5C, D). For 43/47 independent events tested, the 598bp repeat remained at the empty donor site (Figure 5C, lanes 1–5). For 2/47 events it was lost from the donor site. Southern blot on F2 and sequencing through the ET500 insertion site on F3 (LL at nga8) confirmed the absence of Ds and the 598 bp repeat in these two cases. For 1/47 events, Southern analysis was consistent with a move of the repeat to another chromosomal position, not necessarily adjacent to a reinserted Ds (Figure 5D, lanes 1, 4). For the remaining one excision, the fate of the fragment was undetermined because results of Southern analysis and PCR were inconclusive.
Figure 5. Excision of Ds with atypical flanking DNA.
(A) Configuration of Ds in ET500 parental Ds element with NPTII, GUS, 5′ and 3′ TIRs. The 598bp inverted repeat is immediately 3′ of the Ds 3′ TIR. The duplicated target half sites (gray boxes) flank the 5′ TIR (typical) and the 598bp inverted repeat (atypical). (B) Excision of Ds yields broken ends that are adjacent to the leftward duplicated target at the 5′ end of Ds and to the 598bp inverted repeat at the 3′ end of Ds, and is predicted to destroy a SacI site. (C) Southern blot analysis of plant DNA digested with SacI, confirming Ds excision. Upper panel (labelled GUS) was probed with GUS sequences, stripped and reprobed with one-half of the 598bp inverted repeat sequence (middle panel, labelled 598bp). Lower portion of blot was cut out and probed with DCL1 cDNA sequences as a loading control (lower panel, labelled DCL1). Lanes 1–5 contained DNA from F2 lines with empty donor sites; lane 6, ET500 parent. The up-shift in band size between ET500 parent and F2 progeny plants in the middle panel is because of destruction of a Sac1 site due to excision. Lanes 1, 2, 4, and 5; F2 progeny had lost Ds from the genome, but retained the 598bp fragment. Lane 3; F2 progeny had a reinsertion of Ds but retained the 598bp fragment at the original locus. (D) Southern blot analysis of SacI digested DNA showing movement of the 598bp segment. Lanes 1 and 2 had F2 DNA from two independent F1 lines, and lane 3 had ET500 parental DNA. Probes were as labelled for panel (C). Lane 1 shows an F1 line in which the Ds and the 598bp inverted repeat had a change of position. Lane 2 shows a line that had lost Ds but not the 598bp inverted repeat sequence from the original location. Hybridization signals expected from the original location are identified with triangles; signals due to sequence alteration are identified with circles (loss of SacI site adjacent to 598 bp repeat), asterisks denote movement of GUS and the 598bp repeat to another location.
We have shown that within the resolution limits of our measurement the excision in Arabidopsis of a Ds element from a homologous chromosome does not provoke interaction of the broken chromosome with its intact homologue, and that the repair mechanism resembles the immunoglobulin V(D)J joining reaction of vertebrates.
Previous experiments showed that at the bronze locus in maize Ac/Ds activity was not associated with homologous recombination . The bronze locus, however, is a recombination hotspot , and thus could have been atypical in its behaviour with respect to Ds excision. Our observations in Arabidopsis together with previous observations in maize at bronze help generalize the conclusion that Ds excision does not frequently influence crossover recombination between homologous chromosome pairs. These results imply that although DNA transposable elements have evolved to provide evolutionary plasticity to the host genome through frequent rearrangements, their excision may seldom affect crossover of flanking arms of homologous chromosomes. We have shown that efficient excision of Ds element can occur despite the absence of canonical target site duplication at the 3′ end of the element. This was also reported for Ac excision from the maize P1-vv varigated pericarp allele that lacks a target site duplication . Finally, based on Southern analysis, the movement of non-Ds inverted repeat elements may be facilitated in cis by the activity of nearby transposable elements. Although we have not provided conclusive molecular evidence for movement of the repeat, our analysis is consistent with the recent finding that endogenous repetitive sequences are involved in a subset of chromosomal rearrangements mediated by Ds excision . A possible mechanism of transposition of the 598 bp repeat could be cryptic transposase binding sites within this inverted repeat. When high-affinity transposase binds to the Ds TIRs, binding of further transposase molecules to nearby cryptic sites may be facilitated. Alternatively, DNA may occasionally slide through the bound transposase whereupon the enzyme makes a cleavage at a distance from the binding site (Fig 4). These interpretations suggest that the formation as well as further transposition of novel inverted repeat sequences are facilitated by existing transposable elements, and could contribute towards the evolution of new mobile elements. The alternative explanation, a move of the 598 bp repeat during a spontaneous rearrangement or recombination at the ET500 locus, not associated with Ds excision and repair, seems unlikely because the control cross ET500×wild type No-0 showed no such instability. More likely are the remaining two explanations: a transposition of the 598 bp repeat to a new chromosomal location, or a transposition during which the Ds and 598 bp repeat were rearranged within the ET500 locus. We suggest that the 598 bp repeat is mobile because we observed a loss of this repeat from the genome in association with two independent Ds transpositions.
In further support of the existence of cryptic binding sites within the 598 bp repeat, motifs were identified based on the results of Becker and Kunze  describing Ac transposase binding sites as ACG, TCG and A/TCGG in the 250 bp subterminal ends inside the 11-bp terminal IRs. We found these motifs 25 times in the 598 bp repeat. Consistent with previous observations  these motifs are present on both strands in direct and inverted repeat orientations. The number of motifs within the 598 bp repeat is comparable to that in Ac ends, namely, 25 in Ac 5′ and 20 in Ac 3′ TIRs, respectively. Although the motifs in the 598 bp inverted repeat are outside the terminal IR of the Ds element, flexibility of the transposase in DNA binding  may allow the transposase to 'slide' and recognize these cryptic sites while still requiring the TIRs of the Ds element in cis for transposition. The presence of transposase binding sites within DNA, however, is necessary but not sufficient for transposition .
The observed lack of detectable crossover enhancement by a pioneering transposable element in a foreign genome and the formation of novel repeated DNA sequences as a consequence of transposon movement have implications on the role of transposable elements in genomic sculpting. While we did not find any evidence of correlated crossover exchange at the site of excision, the influence of transposable elements as agents causing rare crossovers cannot be eliminated. Nevertheless, it appears that movements of transposons are unlikely to be the most important direct agent for causing somatic or germinal crossovers. On the other hand, the production of novel repeated elements might indirectly create the possibility of intra-chromosomal or inter-chromosomal crossover by homologous recombination in subsequent generations.
A consideration of the conserved and unique aspects of Ds footprints in maize , , , Arabidopsis [23; this work] and yeast ,  suggest that binding of the transposase molecules at Ds TIRs may be sufficient for accurate excision of Ds but that the mechanism of repair of broken ends at the donor site is dependent on host factors. In maize and tobacco, repair occurs mostly by an error prone repair synthesis followed by end joining. In Arabidopsis and yeast, the major footprints are consistent with hairpin formation, hairpin opening and microhomology mediated end joining. One wonders whether by manipulating host factors in plants might the ends be forced to repair by homologous recombination as normally seen with P-element transposition in Drosophila , .
Arabidopsis thaliana strain ET500 (La-er) was the source of Ds insertion element , and strain NaeAc (No-0) was the source of Ac transposase . Conditions of growth and propagation were as described before .
ET500 and NaeAc putative homozygous insertion lines were selfed for two generations. In the second generation ET500 F2 segregated 100% (42/42) kanamycin resistant seedlings, and has GUS+ leaves with 100% penetrance and expressivity throughout the life cycle. In addition, a ring of cells on the ovary receptacle expressed GUS. The GUS transgene segregated as a dominant marker in the cross ET500×wild type Nossen: 31/42 of F2 progeny were GUS+, indicating a single dominant GUS transgene (P<0.005 for heterozygosity). Similarly, in the second generation of NaeAc selfing, 14/14 F2 plants were kanamycin resistant and tested positive for the transposase gene by PCR analysis. Additionally all 53 F1 plants of ET500×NaeAc scored positive for the Ac gene by PCR analysis. Sequencing of genomic DNA 301 bp to the left of the Ds insertion site and 341 bp to the right of the inverted repeat sequence showed 100% sequence homology between the NaeAc and Landsberg erecta parental lines.
5′TAACAAGAATGAGATATAAAGTATCACCTTCTCATCATGAG3′). For PCR of excision sites in plants that lost both the Ds and ocs3′-RB (Primers: T6136, 5′CCATTCGCCCTATAGTGAGTCG3′; and T6409-2, 5′TGCCAGTCAGCATCATCACACC3′), the reaction condition was 36 cycles of 30s (96°C)—30s (63°C)—90s (72°C). PCR of the ocs3′-RB inverted repeat flanked by a portion of the 3′ Ds and plant DNA was with 0.25 µM each of primers Ds3-2E (5′CGTTACCGGTATATCCCGTTTTCG3′) and 58179E (5′CAATGCTCTCTGCCTTCGCCGTTAAAG3′) for 36 cycles of 30s (96°C)—30s (61.7°C)—120s (72°C).
PCR of the 598 bp inverted repeat flanked by 3′Ds and plant genomic DNA was performed in a reaction volume of 50 µl with 1M betaine (Sigma) and 6.25 units Taq DNA polymerase (Qiagen), with 0.25 µM each of primers Ds3-2E and 52879E. The reaction was a 'hot start' with 1 cycle of denaturation at 95°C, then 36 cycles of 30 s (96°C)—30s (61.7°C)—2min (72°C).
Thermal asymmetric interlaced (TAIL) PCR  for DNA genomic flanking the 5′ Ds in ET500 was accomplished with primers: Ds5-1, Ds5-2, Ds5-3 and AD3, with secondary and tertiary cycles as described . DNA on the 3′ flank of Ds was cloned by single-stranded PCR base on the "5′ Race System for Rapid Amplification of cDNA Ends, Version 2.0" (GibcoBRL), with modifications as follows: single-stranded PCR of 1 µl of ET500 genomic DNA with Ds3-1 and Taq Polymerase High Fidelity as follows: 1 cycle 94°C for 2 min; 40 cycles of 15 sec at 94°C, 15 sec annealing at 55°C and 1 min extension at 68°C; PCR of C-tailed DNA, Ds3-2 with 20 sec denaturation at 94°C, 15 sec annealing at 50°C and 3 min polymerization at 68°C; 40 cycles; nested PCR with CUDs3-3A, 5′CAUCAUCAUCAUGGTAGAGGTATTTTACCGACC3′, 94°C hot start 20 sec at 94°C, 15 sec at 55°C and 3 min at72°C, 40 cycles. Polymerization for the 40th cycle was extended from 3 to 7 min.DNA fragments cloned from the 3′ flank representing one-half of the inverted repeat were used probing Southern blots. PCR primers for probes were T6136 and T6409-2. Primer T6136 corresponds to 22 bases immediately (3′) of the SacI site, within the inverted repeat, as shown in Fig. 1 (The 5′ end of primer T6409-2 is complementary to base 6409 on the ET500 parental T-DNA vector; Genbank accession number M35007.1), and its 3′ end is complementary to base 6378 on the same T-DNA sequence. The 5′ end crosses the point of inversion by three bases (Fig. 1). Sequencing of the NaeAc and wt Landsberg erecta parental genomic DNA flanking the ET500 insertion site for comparison of homology was accomplished with primers 318E and 58179E.
Sequencing reactions were as stated in the ABI Prism BigDye Terminator cycle Sequencing Ready Reaction Kit (Applied Biosystems), with 16 pmoles of primer and 6ul Big dye Terminator Ready Reaction Mix (Applied Biosystems) for a 20 ul reaction. Sequencing primers and PCR primer sequences were the same. To sequence the 598 bp inverted repeat with flanking Ds and plant DNA, the reaction was 40 µl with 32 pmoles Ds3Sac, 5′CCGTTTTCATCCCTAGAGCTCCAATTCGC3′, 1M betaine (Sigma) 16 µl Big Dye Terminator Ready Reaction Mix, 1M betaine as follows: (98°C)-3min, then 25 cycles of 30s (96°C)—4min (64.0°C).
We thank Drs. Peter Gibbs, Teresa Golden, Martin Gorovsky, David Hinkle, Christopher Lawrence, Stephen Schauer, Samuel Schlagman, and Clifford Weil for discussion and advice, Dr. Nina Fedoroff for the Ac strain, and Wendy B. Gagliano for technical assistance.
Conceived and designed the experiments: AR UG ML. Performed the experiments: ML LS. Analyzed the data: AR ML. Contributed reagents/materials/analysis tools: AR UG. Wrote the paper: AR ML. Other: Edited the paper: UG. In the PhD committee of Marybeth Langer: UG. Undergraduate researcher who worked under Marybeth Langer's supervision: LS. Advised Marybeth Langer's PhD research work: AR.
1. Kazazian HH Jr (2004) Mobile elements: drivers of genome evolution. Science 303: 1626–1632.
2. Ros F, Kunze R (2001) Regulation of activator/dissociation transposition by replication and DNA methylation. Genetics 157: 1723–1733.
3. Rinehart TA, Dean C, Weil CF (1997) Comparative analysis of non-random DNA repair following Ac transposition excision in maize and Arabidopsis. Plant J 12: 1419–1427.
4. Weil CF, Kunze R (2000) Transposition of maize Ac/Ds transposable elements in the yeast Saccharomyces cerevisiae. Nat Genet 26: 187–190.
5. Rommens CM, van der Biezen EA, Ouwerkerk PB, Nijkamp HJ, Hille J (1991) Ac-induced disruption of the double Ds structure in tomato. Mol Gen Genet 228: 453–458.
6. Van der Biezen EA, Brandwagt BF, van Leeuwen W, Nijkamp HJ, Hille J (1996) Identification and isolation of the FEEBLY gene from tomato by transposon tagging. Mol Gen Genet. 251: 267–280.
7. Eichler EE, Sankoff D (2003) Structural dynamics of eukaryotic chromosome evolution. Science 301: 793–797.
8. McClintock B (1953) Mutations in maize. Carnegie Inst Wash Ybk 52: 227–237.
9. Fradkin CMW, Brink RA (1956) Crossing over in maize in the presence of the transposable factors Activator and Modulator. Genetics 41: 901–906.
10. Fischer SE, Wienholds E, Plasterk RH (2003) Continuous exchange of sequence information between dispersed Tc1 transposons in the Caenorhabditis elegans genome. Genetics 164: 127–134.
11. Xiao YL, Li X, Peterson T (2000) Ac insertion site affects the frequency of transposon-induced homologous recombination at the maize p1 locus. Genetics 156: 2007–2017.
12. Wright SI, Agrawal N, Bureau TE (2003) Effects of recombination rate and gene density on transposable element distributions in Arabidopsis thaliana. Genome Res 13: 1897–1903.
13. Fu H, Zheng Z, Dooner HK (2002) Recombination rates between adjacent genic and retrotransposon regions in maize vary by 2 orders of magnitude. Proc Natl Acad Sci USA 99: 1082–1087.
14. Yao H, Zhou Q, Li J, Smith H, Yandeau M, et al. (2002) Molecular characterization of meiotic recombination across the 140-kb multigenic a1-sh2 interval of maize. Proc Natl Acad Sci USA 99: 5763–5765.
15. Coen ES, Carpenter R, Martin C (1986) Transposable elements generate novel spatial patterns of gene expression in Antirrhinum majus. Cell 47: 285–96.
16. Bai L, Singh M, Pitt L, Sweeney M, Brutnell TP (2007) Generating novel allelic variation through Activator insertional mutagenesis in maize. Genetics 175: 981–992.
17. Roth DB, Zhu C, Gellert M (1993) Characterization of broken DNA molecules associated with V(D)J recombination. Proc Natl Acad Sci USA 90: 10788–10792.
18. McBlane F, van Gent D, Ramsden D, Romeo C, Cuomo C, et al. (1995) Cleavage at a V(D)J. recombination signal requires only RAG1 and RAG2 proteins and occurs in two steps. Cell 83: 387–395.
19. Sundaresan V, Springer P, Volpe T, Haward S, Jones JD, et al. (1995) Patterns of gene action in plant development revealed by enhancer trap and gene trap transposable elements. Genes Dev 9: 1797–1810.
20. Zhou L, Mitra R, Atkinson PW, Hickman AB, Dyda F, et al. (2004) Transposition of hAT elements links transposable elements and V(D)J recombination. Nature 432: 995–1001.
21. Yu J, Marshall K, Yamaguchi M, Haber JE, Weil CF (2004) Microhomology-dependent end joining and repair of transposon-induced DNA hairpins by host factors in Saccharomyces cerevisiae. Mol Cell Biol 24: 1351–1364.
22. Scott L, LaFoe D, Weil CF (1996) Adjacent sequences influence DNA repair accompanying transposon excision in maize. Genetics 142: 237–246.
23. Gorbunova V, Levy AA (2000) Analysis of extrachromosomal Ac/Ds transposable elements. Genetics 155: 349–359.
24. Xiao YL, Peterson T (2002) Ac transposition is impaired by a small terminal deletion. Mol Genet Genomics 266: 720–731.
25. Conrad LJ, Brutnell TP (2005) Ac-immobilized, a stable source of Activator transposase that mediates sporophytic and gametophytic excision of Dissociation elements in maize. Genetics 171: 1999–2012.
26. Szostak JW, Orr-Weaver TL, Rothstein RJ, Stahl FW (1983) The double-strand-break repair model for recombination. Cell 33: 25–35.
27. Dooner HK, Martínez-Férez IM (1997) Germinal excisions of the maize transposon activator do not stimulate meiotic recombination or homology-dependent repair at the bz locus. Genetics 147: 1923–1932.
28. Dooner HK, Martínez-Férez IM (1997a) Recombination occurs uniformly within the bronze gene, a meiotic recombination hotspot in the maize genome. Plant Cell 9: 1633–1646.
29. Page DR, Koehler C, da Costa Nunes JA, Baroux C, Moore JM, et al. (2004) Intrachromosomal excision of a hybrid Ds element induces large genomic deletions in Arabidopsis. Proc Natl Acad Sci USA 101: 2969–2969.
30. Becker HA, Kunze R (1997) Maize Activator transposase has a bipartite DNA binding domain that recognizes subterminal sequences and terminal inverted repeats. Mol Gen Genet 254: 219–230.
31. Saedler H, Nevers P (1985) Transposition in plants: a molecular model. EMBO J 4: 585–590.
32. Yan X, Martínez-Férez IM, Kavchok S, Dooner HK (1999) Origination of Ds elements from Ac elements in maize: evidence for rare repair synthesis at the site of Ac excision. Genetics 152: 1733–1740.
33. Engels WR, Johnson-Schlitz DM, Eggleston WB, Sved J (1990) High-frequency P element loss in Drosophila is homolog dependent. Cell 62: 515–523.
34. Gloor GB, Nassif NA, Johnson-Schlitz DM, Preston CR, Engels WR (1999) Targeted gene replacement in Drosophila via P element-induced gap repair. Science 253: 1110–1117.
35. Fedoroff NV, Smith DL (1993) A versatile system for detecting transposition in Arabidopsis. Plant J 3: 273–289.
36. Golden T, Schauer SE, Lang JD, Pien S, Mushegian A, et al. (2002) SHORT INTEGUMENTS1 / SUSPENSOR1 / CARPEL FACTORY, a Dicer homologue, is a maternal-effect gene required for embryo development in Arabidopsis. Plant Physiol 130: 808–822.
37. Ray A, Lang JD, Golden T, Ray S (1996) SHORT INTEGUMENTS1 (SIN1), a gene required for ovule development in Arabidopsis, also controls flowering time. Development 122: 2631–2638.
38. Liu YG, Mitsukawa N, Oosumi T, Whittier RA (1995) Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junction by thermal asymmetric interlaced PCR. Plant J 8: 457–463.
39. Grossniklaus U, Vielle-Calzada JP, Hoeppner MA, Gagliano WB (1998) Maternal control of embryogenesis by MEDEA, a Polycomb group gene in Arabidopsis. Science 280: 446–450.


For Over 21 Years , the experts at Flooring Design Center have been providing quality, name-brand products at great prices to customers in Northern Virginia, DC and Maryland. Flooring Design Center provides services to residential and business customers with a wide selection of products suitable for homes, small businesses, organizations and commercial applications. Our line of quality, name-brand products includes: Carpet, Hardwood, Laminate, Ceramic, Vinyl Flooring, Complete Bathroom Fixtures, Cabinets and Granite Counter-tops.
Our company provides shop-at-home convenience, professional installation and quality products at great prices. With over 30,000 satisfied customers, flooring design center is dedicated to customer satisfaction.
With a 5,000 square foot local warehouse, Flooring Design Center stocks hundreds of products available for immediate delivery & installation by our in-house Installers. As a local company, Flooring Design Center employs Professional Staff & Installers, with an average of 15 years of experience in the Industry.
For our Commercial Customers, Flooring Design Center' dedicated Project Managers and our capability to stock pre-selected materials will ensure high-quality work with on-time delivery & completion on every project. Flooring Design Center, a locally-owned company with the experience, knowledge and facilities to deliver quality every time.


Cedar shake enjoys continued market demand even though more durable and low maintenance roofing materials, such as metal and vinyl, are available. Cedar shake has a unique aesthetic appeal, and its brown shade can enhance the appearance of rustic-style buildings. On the other hand, even the highest quality cedar shake roofing material requires more TLC than other roofing materials.
Shell Busey, in a recent article that was featured on Canada.com, advises caution on the kind of TLC you give your biodegradable cedar shake roof. Roof cleaning products and methods remain largely unregulated, and the use of improper cleaning products and methods could damage your cedar shake roofing. In fact, even the act of pressure washing could inflict irreversible damage.
Cedar wood is renowned for its endurance and can survive for centuries. The natural oils found in cedar trees enables it to live for as long as a thousand years. For roof cleaning, however, the Cedar Shake and Shingle Bureau (CSSB) recommends topical treatments that come with a material safety data sheet (MSDS) that lists product ingredients and safety precautions, as well as certifications.
Do not use a topical treatment product that claims to be fire-retardant or have a 10-year effectiveness. Also avoid sealants, waterproofers or plasticizers, or products that contain unfortified linseed oil, diesel fuel or crankcase oil. Topical solutions such as latex, butyl or silicon, "seal" or coat the surface of the cedar preventing it from "breathing". Anything used as a topical treatment should be [labeled] as a cedar roof treatment product or have a manufacturer's letter stating that it's appropriate for treating cedar roofs.
For quality wood roofing supplies, home builders, contractors, homeowners, and resellers should get their cedar shingles and shakes from renowned suppliers like WoodRoof.com. High quality cedar shingles and shakes are biodegradable, wind resistant, light weight, impact resistant, and of course, durable.