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.
出处
《生物技术通讯》
CAS
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)
