文档介绍：重组酶Cre的逆转录病毒载体的构建及鉴定
作者:陆群吴补领张洪伟王冀姝韩骅余擎苏凌云
【关键词】逆转录病毒
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-,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 . .
Keywords:bination,ic;gene knockout;retro-virus;ic vector
Abstract:AIM To construct binase Cre expressing retrovirus expression vector and infect the mouse host cell in Cre gene was inserted into retrovirus expression vector pMx-IRES-GFP and the packing cell line was transfected to p
roduce retrovirus particles infected NIH-3T3cells in vitro,and the virus titer was determined by green lusaferase binase Cre expressing retrovirus expression vector pMx-IRES-GFP-Cre was constructed essfully,and the packing cell line was able to produce retrovirus virus titer was105 cfu mL-1 .CONCLUSION binase Cre would be expressed by retrovirus expression system,and the retrovirus particles can infect the host cells system will be helpful in studying the function of specific gene.
0 引言
Cre是P1噬菌体表达的重组酶,(floxed基因),都可以被Cre重组酶剪切[1] .因此Cre-LoxP系统目前成为一种普遍应用的条件性剔除基因的方法;为了研究Cre重组酶在体外细胞水平对LoxP位点的识