重组酶Cre的逆转录病毒载体的构建及鉴定

Construction and identification of recombinase Cre expressing retrovirus expression vector

目的 构建表达重组酶 Cre的逆转录病毒载体并进行初步的体外感染研究 .方法 将 Cre基因片段插入到逆转录病毒载体 p Mx- IRES- GFP中 ,转染包装细胞系获得表达Cre的病毒颗粒 ,体外感染 NIH- 3T3细胞 ,通过绿色荧光蛋白GFP测定病毒滴度 .结果 构建了表达 Cre的逆转录病毒载体 p Mx- IRES- GFP- Cre.通过包装细胞系得到的含有病毒颗粒的培养上清 ,这种培养上清具有体外感染细胞的能力 ,病毒滴度为 10 5cfu·m L- 1 .结论 用逆转录病毒体系表达重组酶Cre并能有效地进行体外感染 .

AIM To construct recombinase Cre expressing retrovirus expression vector and infect the mouse host cell in vitro . METHODS Cre gene was inserted into retrovirusexpression vector pMx IRES GFP and the packing cell line was transfected to produce retrovirus particles. Retrovirus particles infected NIH 3T3 cells in vitro , and the virus titer was determined by green lusaferase protein. RESULTS Recombinase Cre expressing retrovirus expression vector pMx IRES GFP Cre was constructed successfully, and the packing cell line was able to produce retrovirus particles. The virus titer was 10 5 cfu·mL -1 . CONCLUSION Recombinase Cre would be expressed by retrovirus expression system, and the retrovirus particles can infect the host cells effectively. This system will be helpful in studying the function of specific gene.