HIV-1跨膜蛋白gp41的截短及表达

Truncation and expression of HIV-1 trans-membrane protein gp41

将HIV 1跨膜蛋白gp4 1进行截短 ,在大肠杆菌中进行表达并纯化。PCR扩增 gp4 1的部分编码基因 ,回收的PCR产物纯化后克隆到连接载体 pGEM T上 ,然后用EcoRⅠ和Sa1Ⅰ切下目的基因 ,并构建到表达载体pGEX 4T3上 ,导入宿主细胞BL2 1(DE3) ,用IPTG诱导表达 ,表达产物用亲和层析进行纯化并作相应鉴定。截短的HIV 1跨膜蛋白 gp4 1能直接在大肠杆菌内进行表达 ,利用亲和层析能方便地将目的蛋白进行纯化 。

To truncate HIV 1 trans membrane protein gp41 and express the truncated gp41(tgp41) in E.coli. The trans membrane protein gp41 was truncated by PCR, the purified PCR products was ligated to cloning vector pGEM T, then the pGEM T tgp41 was digested with EcoRⅠand Sal Ⅰ , the tgp41 fragment was isolated and inserted to the corresponding restriction site on expression vector pGEX 4T 3. The recombinant plasmid pGEX 4T 3 tgp41 was transformed to host BL21(DE3), then expressed by adding IPTG. The truncated gp41 can be expressed in E.coli. and easy to be purified with affinity chromatography. It laid a foundation of further development of trans membrane protein.