PCR法直接筛选重组阳性克隆  

Screening for recombinant clones by colony polymerase chain reaction

作　　者：姜宏[1] 李麓芸[2] 卢光琇[2] 

机构地区：[1]解放军105医院生殖中心,合肥230031 [2]中南大学湘雅医学院生殖工程研究室

出　　处：《安徽医学》2005年第2期89-90,共2页Anhui Medical Journal

摘　　要：目的　建立一个简便、可靠的重组阳性克隆的筛选方法。方法　应用扩增小鼠生精细胞凋亡相关基因时使用的引物 ,以基因重组扩增后得到的菌落为模板 ,直接进行PCR扩增。结果　在筛选的阳性菌落中可扩增到与小鼠凋亡相关基因片段大小一致的阳性条带 ;以阳性克隆提取质粒进一步进行PCR和酶切鉴定及序列分析 ,证明结果正确。结论　菌落PCR是一种简便、快速、可靠、有效的筛选重组阳性克隆的方法。Objective To develope a reliable method for rapidly screening positive clones of recombinant genes.Methods The recombinant colonies were transfered into the PCR reaction mixture. The PCR was carried out by using primers for amplificating genes related to apoptosis in spermatogenic cells of mouse.Results The same size positive strips as the cDNA fragments related to apoptosis in spermatogenic cells of mouse were visible in the colony PCR of the screened positive clones. The positive colony were further confirmed by the PCR with plasmid DNA and digestions as well as DNA sequencing. Conclusion Colony PCR is a simple, efficient and reliable technique for screening positive clones of recombinant genes.

关 键 词：PCR扩增 小鼠 PCR法 菌落PCR 筛选 凋亡相关基因 生精细胞凋亡 阳性克隆 酶切 质粒 

分 类 号：R346[医药卫生—基础医学]