t-PA突变体山羊乳腺表达载体构建及转染细胞的核移植试验  

The construction of goat mammary gland expressed vector pGBCt-PAm and the study of nuclear transfer

作　　者：杨正田[1] 兰国成[2] 沈伟[1] 吴晓洁[1] 徐秀英[1] 谭景和[2] 邓继先[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071 [2]山东农业大学动物科技学院,泰安271018

出　　处：《生物技术通讯》2004年第5期433-436,共4页Letters in Biotechnology

基　　金：国家高技术研究发展计划重大专项(2002AA206621)

摘　　要：构建了t-PA突变体山羊乳腺特异表达载体pGBCt-PAm,包括长4.1kb的山羊β-酪蛋白启动子以及启动子前2.4kb的鸡β-珠蛋白绝缘子序列;并检测了表达载体在家兔乳腺的瞬间表达,验证了载体的有效性。通过电击和脂质体2种转染方式转染了山羊胎儿成纤维细胞(GFFC),经过筛选检测,得到了7株能大量扩增的PCR阳性细胞株。其中的2株(M9,M10)已经进行了首次细胞核移植和胚胎移植试验,2只受体羊2个情期未见返情。The mammary gland expression vector pGBCt-PAm including 4.1 kb goat β-casein promoter and 2.4 kb chicken β-globin insulator in front of the promoter was constructed. The availability of the vector was testified through temporal expression in the mammary gland of the female rabbit. The goat fetal fibroblast cells were transfected by pGBCt-PAm with electroporation or lipidosome, and after the identification of PCR, 7 lines of progenitive cells that examined positive were got, the two(M9,M10)of which had been used for the somatic nuclear transfer for the first time. After embryo transfer, 2 recipient goats have not been seen oestrous within 2 oestrum.

关 键 词：t-PA 乳腺表达 转染细胞 首次 羊胎 阳性细胞 脂质体 Β-酪蛋白 核移植 突变体 

分 类 号：Q786[生物学—分子生物学] Q78