1 5 CHO dNTP dhfr- 25/0 و High Glucose, GlutaMAX Gibco Pfu 75/10 °C94 7 PCR 15 °C94 1 و Gibco سپس و °C 50 30 DMEM °C 72 و °C 72 10 و DMEM, High Glucose, GlutaMAX + 1/0 mM hypoxanthine, 016/0 mM thymidine و EcoRI BglII 002/0 mM Methotrexate+10% FBS+P/S 1% pS-IFN Kozak Invitrogen و Lipofectamine® LTX و PLUS™ Reagent و و MBI fermentas و EcoRI و Metabion BglII TangoTM 80-50 % داد FermentasMBI lµ و 12 lµ 1200 500 و ng DNA 3 و lµ سپس 12 (lµ1 PLUS™ Reagent DNA وBglII u/µl10 EcoRI DNA PLUS™ 1 DNA PCR 5 Reagent 5/0 20 1 و lµ 30 Lipofectamine® (lµ5/2 LTX و 30 lµ 100 و 37 5% و CO2 Megaprimer Qiagen 206 UTR ARE´3 2/0 18 10 و 550 IFNβ T4 DNA Ligase MBI pS-IFN Kozak Fermentas 100 80 µl1 و µl X10 و و µl20 16 PCR سپس °C4 و pS-IFN Kozak IFNβ E. coli و سپس و 18 UTR ARE´3 pS-IFNΔARE DNA coli E. E. coli E. coli CHO dftr- LBA و RNA و 1 37 CHO (CHO NCBI Code: C590) dftr-
dNTP 1 5 Pfu 25/0 و 75/10 PCR °C94 7 و سپس 15 °C94 1 °C 50 30 °C 72 و °C 72 10 EcoRI و BglII pS-IFN Kozak و EcoRI و MBI fermentas و BglII Metabion TangoTM FermentasMBI و 3 و EcoRI وBglII u/µl10 1 DNA PCR 5/0 1 و 20 Qiagen 2/0 و 10 T4 DNA Ligase MBI Fermentas 80 µl X10 و µl1 و µl20 سپس 16 °C4 و E. coli و سپس DNA E. coli CHO dftr- RNA و CHO (CHO dftr- NCBI Code: C590) CHO dhfr- High Glucose, GlutaMAX Gibco و Gibco DMEM و DMEM, High Glucose, GlutaMAX + 1/0 mM hypoxanthine, 016/0 mM thymidine 002/0 mM Methotrexate+10% FBS+P/S 1% Invitrogen Lipofectamine® LTX و PLUS™ Reagent و و 80-50 % داد lµ 100 12 lµ 1200 و ng 500 DNA سپس lµ 12 PLUS™ Reagent (lµ1 DNA DNA PLUS™ Reagent 5 lµ 30 Lipofectamine® LTX (lµ5/2 و 30 lµ 100 و 37 و CO2 5% Megaprimer 206 UTR ARE´3 18 550 IFNβ pS-IFN Kozak و PCR pS-IFN Kozak IFNβ و UTR ARE´3 18 pS-IFNΔARE E. coli E. coli LBA و 1 37