外源基因转染山羊体细胞的整合效率研究  

Study on Integration Efficiency of Foreign Genes Transfecting to Goat Somatic Cells

作　　者：蔡琳琳[1,2] 蔡勤[1,2] 许淼[1,2] 黄英[1,2] 

机构地区：[1]上海市儿童医院,上海医学遗传研究所 [2]卫生部医学胚胎分子生物学重点实验室暨上海市生殖与胚胎工程重点实验室,中国上海200040

出　　处：《生命科学研究》2013年第1期32-35,共4页Life Science Research

基　　金：国家科技重大专项资助项目(2009ZX08007-003B)

摘　　要：为了探讨影响山羊体细胞中外源基因整合效率的因素,采集胎羊和小羊皮肤并制备成纤维细胞,采用"Lipofectamine LTX"脂质体转染试剂包埋外源基因,分别导入上述细胞中,比较同一种外源基因载体(人凝血因子Ⅸ)转染至不同个体和类型的羊细胞,以及不同载体(人凝血因子Ⅸ、人转铁蛋白和牛催乳素)转染至同一个体羊细胞的整合效率的差异.通过药物筛选得到单细胞簇,经定量PCR计算外源基因的整合效率.同一种外源目的基因载体转染小羊成纤维细胞的整合效率显著高于胎羊成纤维细胞(P<0.05);而不同目的基因载体转染同一个体羊细胞,其细胞的整合效率与转染载体的片段大小有关.由此可知,小羊成纤维细胞外源基因整合频率优于胎羊细胞,且载体片段的大小影响外源基因转染的整合频率.To investigate the impacts of foreign DNAs in goat somatic cells on the integration efficiencies, the skins were collected from fetal and young goats and the fibroblasts were then cultured. The foreign gene vectors were transfected into the goat fibroblasts using＂Lipofectamine LTX＂, and the integrated frequencies were com- pared between the Cells derived from different types of goats transfected with the same foreign vector （hFIX） as well as between the cells derived from the same goat but transfected with three different vectors （hFIX, hTF or bPRL）. The monocloned cell clusters were obtained with the drug selection and the integrated frequency was calculated with qPCR. The integration efficiency of the young goat fibroblasts was higher than the fetal cells （P〈0.05）. In the same cells transfected with different foreign genes, the integration frequencies were relevant with the length of the vectors. In conclusion, the integrated efficiency in young goat fibroblasts was better than in the fetal cells for transfected the same foreign gene. In addition, the sizes of the DNA fragments transfected had the effect on integration efficiency.

关 键 词：基因转染 山羊体细胞 整合效率 体细胞核移植技术 

分 类 号：Q812[生物学—生物工程]