脂质体转染胚盘细胞制备携pEGFP-C2基因的转基因鹌鹑  

Generation of pEGFP-C2 expression transgenic quail by liposome transfection blastoderm cells

作　　者：李艳[1] 王小义[1] 李青峰[1] 赵宗胜[1] 

机构地区：[1]石河子大学动物科技学院,新疆石河子832003

出　　处：《中国兽医学报》2014年第3期442-445,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30660125)

摘　　要：利用脂质体转染第X期胚盘细胞的方法将外源质粒pEGFP-C2经脂质体包裹后注射到鹌鹑种蛋X期胚盘下腔,处理种蛋110枚,封口后孵化。出壳9只G0代,出壳率为8.18%。经检验6只鹌鹑中有4只为阳性,阳性率为66.67%。对成年的4只转基因鹌鹑中的1只内脏组织进行PCR分析以及切片荧光检测,证实外源基因在G0代成年鹌鹑组织中成功表达。将3只成年转基因鹌鹑分别与野生型鹌鹑进行杂交,得到G1代阳性率分别为33.96%、20.59%、10%。再将3只成年转基因公母鹌鹑之间进行杂交,得到G1代阳性率分别为10.87%、42.86%。证实利用脂质体转染胚盘细胞制备转基因鹌鹑是可以将外源基因整合到G0代生殖系中,且外源基因能遗传给后代。To explore the possibility of transgenic quail generated by liposome transfection blasto- derm cells, foreign plasmid pEGFP-C2 was injected into the subgerminal cavity of the fresh fertil- ized eggs before incubation,out of the 110 microinjected eggs,9 Go chicks were hatched （8.18%）, revealing 4 positives（66.67 %）of the total. Using one of four Go generation mature quail tissue for DNA extraction and PCR analysis,and tissue sections for fluorescent detection, the expression of foreign gene in Go generation quail tissue was confirmed. Three adult transgenic quail passed the transgene onto its offspring when mated with wild-type quail, the positive ratio was 33. 96%, 20.5 9 %, and 10 %, respectively. Then crossing among three adult transgenic quails, the G1 genera- tion positive rate was 10.87% and 42.86%. Confirming the use of liposomal transfection the blas- toderm cell for generatian of transgenic quail can integrate exogenous gene to Go generation repro- ductive system,and the foreign gene to their offspring.

关 键 词：脂质体转染 成年转基因鹌鹑 荧光检测 阳性率 

分 类 号：S852.2[农业科学—基础兽医学]