脂质体转染胚盘细胞将外源质粒pEGFP-C2导入鹌鹑胚的研究  

Introduction of foreign plasmid pEGFP-C2 into the quail embryo via liposome transfection blastoderm cells

作　　者：李艳[1] 李洪涛[1] 段丹丹[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》2013年第4期592-596,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30660125)

摘　　要：运用脂质体转染第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,孵化种蛋。提取孵化6d的鹌鹑胚基因组DNA,进行PCR检测,嵌合体阳性率为84%,证实外源基因在胚胎中的表达。对孵化至出壳的G0代鹌鹑组织进行DNA提取并PCR分析,以及组织切片荧光检测,证实外源基因在G0代鹌鹑组织中成功表达。结果表明脂质体转染胚盘细胞是一种制备转基因鹌鹑行之有效的方法。In order to identify the availability of liposome transfection of foreign DNA as an effective way in making transgenic quail,foreign plasmid pEGFP-C2 was injected into the subgerminal cavity of the fresh fertilized eggs before incubation,and the embryos of 6 day-old were collected to identify the distribution of the exogenous DNA, The positive ratio was 84 %, the exogenous gene was expressed in embryos. Using Go generation quail tissue for DNA extraction and PCR analysis, and tissue sections for fluorescent detection,the successful expression of foreign gene in Go generation quail tissue was confirmed. This research results indicates the method is effective to produce transgenic quail.

关 键 词：脂质体转染 胚盘细胞 荧光检测 

分 类 号：Q78[生物学—分子生物学]