牛的3种单个早期胚胎中基因表达的差异

为建立哺乳动物单个早期胚胎中基因表达的分析方法,解决胚胎实验中取材困难的矛盾,本研究以牛体内受精胚(in vivo cultured,IVC)、体外受精(in vitro fertilization,IVF)和体细胞核移植(somatic cell nuclear transfer,SCNT)3种来源的7日龄囊胚作实验材料,通过单个胚胎直接裂解后进行逆转录结合随机引物指数扩增的方法富集相应cDNA.对克隆囊胚cDNA库中随机挑选的15个阳性克隆进行测序和EST分析.并运用特异性的引物,对哺乳动物胚胎发育的3个重要基因Nanog,Oct-4和Sox2的表达进行了扩增和表达检测.结果表明,在哺乳动物胚胎囊胚阶段,这3个维持细胞多能性的相关基因的表达都相当活跃;相对于体内胚和体外受精胚,克隆胚中这3种基因的表达明显偏低.本方法为牛植入前胚胎发育阶段特异性基因的筛选和基因表达模式的研究提供了有效的材料和技术途径.

Different Gene Expression among Early Single Bovine Embryos of Three Kinds

To set up the technique by which cellular gene expression analysis could be done from a single preimplantation embryo of mammalian so as to resolve the challenging problem of accurate research on limited amount of homogeneous and shortage materials in some cases.Three kinds of single 7-day embryos derived from in vivo cultured(IVC),in vitro fertilization(IVF),and somatic cell nu-clear transfer(SCNT) were used as materials.Single embryo was lysed directly in lysis buffer and then mRNA was reverse transcribed...