摘要
为了优化利用体细胞核移植生产转基因牛早期胚胎的体系,以携带绿色荧光蛋白-新霉抗性双标记基因的pMSCV质粒转染的胎牛耳成纤维细胞为供体,以体外成熟的牛卵母细胞为受体构建克隆胚,研究供体细胞的传代次数对转基因胚的影响和重构胚在不同电场条件下(电场强度、直流电脉冲次数)融合效率及其在不同体外培养系统中的发育效果。结果表明:传代5次的细胞做核供体较有利于转基因胚的发育;在电场条件为电场强度1.6 kV/cm、直流脉冲1次的融合率最高;负压单层细胞共培养体系中的转基因囊胚发育率较好,与未转基因体细胞核移植胚的发育相比无显著差异(P>0.05)。
This study was to optimize a system for production of transgenic bovine embryos by nuclear transfer of somatic cells. PMSCV fetal fibroblast cells with green fluorescent protein - mycophenolate resistance marker gene was the donor. The maturation of bovine oocyte in vitro was receptor. The embryo cloning was constructed by the donor and receptor. Under different conditions , such as electric field intensity and DC pulse frequency , the passage of the number of cells of transgenic embryos and integration will be studied. The donor cells of five passages was beneficial to transfer embryo. The rate of the integration was the highest under 1.6 kV / cm of the electric field strength and DC pulse repetition frequency of 1 and negative monolayer co - culture system.
出处
《黑龙江畜牧兽医》
CAS
北大核心
2008年第12期16-19,共4页
Heilongjiang Animal Science And veterinary Medicine
关键词
牛
体细胞核移植
转基因
体外培养
Bovine
somatic cell nuclear transplantation
transgenic
in vitro
