关键词: 重组，遗传;基因剔除;逆转录病毒;遗传载体

    摘 要: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pMx-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pMx-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105 cfu mL-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.为用这一方法在体外研究特异基因对细胞生物学行为的影响奠定基础.

    Construction and identification of recombinase Cre expressing retrovirus expression vector

    LU Qun WU Bu-Ling ZHANG Hong-Wei WANG Ji-Shu Han Hua YU Qing SU Ling-Yun

    1 Department of Oral Conservative，Stomatological College，

    2 Department of Gastrointestinal Surgery，Xijing Hospital，

    3 Department of Genetics and Growth Biology，Faculty of Preclinical Medicine，Fourth Military Medical University，Xi'an710033，China

    Keywords:recombination，genetic;gene knockout;retro-virus;genetic vector

    Abstract:AIM To construct recombinase Cre expressing retrovirus expression vector and infect the mouse host cell in vitro.METHODS Cre gene was inserted into retrovirus expression vector pMx-IRES-GFP and the packing cell line was transfected to produce retrovirus particles.Retrovirus particles infected NIH-3T3cells in vitro ......