Everything about Bacteriophage

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Lambda Phages

The lambda phage, also called Enterobacteria phageλ andcolphageλ, is a type of temperate bacteriophage or bacterial virus that infects the Escherichia coli (E. coli) species of bacteria. The virus may be housed in the genome of its host via lysogeny.

History of Lambda Phage

In 1950, Esther Lederberg, an American microbiologist, was performing experiments on E. coli mixtures. She happened to observe streaks of mixtures of two types of E. coli strains that seemed to have been nibbled on and had viral plaque. One E. coli strain had been treated with ultraviolet light, so the damage was more apparent. It was later determined that this was caused by bacterial viruses, which replicated and spread resulting in cell destruction. The discovery led to the employment of Lambda phage as a model organism in microbial genetics as well as in molecular genetics.

Structure

A lambda phage has a head measuring around 50-60 nanometers in diameter and a flexible tail that is around 150 nanometers long and may contain tail fibers.

The head consists of various proteins and over a thousand protein molecules including X1, X2, B, B*, E, D, and W. The head functions as a capsid that contains its genome, which contains 48,490 base pairs of double-stranded linear DNA. This number also includes 12-base single-stranded parts at its 5’ ends. The single-stranded parts are known as sticky sites and are also called the cos site, which encircles the DNA in the host cytoplasm. Hence, when in circular form, the phage genome is comprised of 48,502 pairs in length. The weight of the genome is estimated to be 32×106 Da, which is around half of the weight of the phage.

The tail has a 135 nanometer-tube that is hollow and contains a conical cap which is around 15 nanometers. The tail’s inner diameter is around 3 nanometers, while on the outside, it is around 9-18 nanometers depending on the knob-like structures that give the tail a rough appearance.

Life cycle

When E. coli is infected with a lambda phage, two cycles may happen: lytic or lysogenic.

The lytic cycle happens when progeny phage particles are produced. The lytic cycle is the more common life cycle that comes after most infections. The first step of this cycle is the attachment of the phage to the host cell, injecting its DNA into the cell. Nucleic acid from the phage is replicated, and the phage’s genes are expressed, allowing the production of phage proteins. The phage proteins are assembled into phage particles, which are released when the host cell undergoes lysis (it breaks down). The lysis is mediated by lysis genes S, R, Rz, and Rz1 which, upon expression, yield proteins that work together to break down the host bacterium’s cell wall. This mode of lambda replication typically yields many phage particles.

The lysogenic cycle, in contrast, does not produce a huge number of progeny phage or break down the host cell. Instead, the λ DNA recombines with its host’s genome to produce a prophage. This typically is the favoured pathway when unfavourable environmental conditions prevent intense replication of the bacterial cells. Like the lytic cycle, the first step of the lysogenic cycle is also the attachment of the phage and the injection of its DNA into the host cell. The phage DNA then integrates with the host chromosome, producing an integrated DNA combination called the prophage DNA. Host cells that carry this DNA are said to be in the lysogenic state. The prophage DNA is replicated along each time the host bacterial cell replicates itself, producing more cells, each with a copy of the prophage DNA. When these cells are exposed to certain chemicals or to ultraviolet light, phage induction happens; the prophage DNA is then cut out of the host genome and proceeds to the lytic cycle.

Applications

The lambda phage has different applications, most of which are related to DNA cloning. This is because lambda phage can be used as a vector for generating recombinant DNA, which are combined DNA sequences that result from using laboratory techniques like molecular cloning to assemble genetic material from several sources. The site-specific recombinase of lambda phage can be used for shuffling cloned DNAs via the gateway cloning system, a molecular biology technique that ensures the effective transfer of DNA fragments between plasmids.

The lambda phage’s ability to mediate genetic recombincation is due to its red operon, which is a functioning unit of genomic DNA that has a cluster of genes controlled by a promoter or a single regulatory signal. This red operon can be expressed to yield the proteins red alpha (or exo), beta, and gamma, which can be used in recombination-mediated genetic engineering, a method commonly employed in bacterial genetics, generation of target vectors, and DNA modification.