【摘要】 基因敲除是建立在同源重组技术基础之上的一种定点修饰基因组的实验方法,其与体细胞核移植技术的成功结合,已经成为一种有效的定点修饰、改造大动物基因组DNA片段的实验策略。朊蛋白是传染性海绵状脑病的致病蛋白。目前,在小鼠和绵羊中已经成功获得朊蛋白基因敲除的动物,但在牛中尚未有成功的报道。文章旨在介绍牛朊蛋白基因敲除无启动子打靶载体的构建过程及技巧,以为将来利用核移植技术生产敲除朊蛋白基因的克隆牛提供依据。更多还原

【Abstract】 Gene knock-out is an approach to introduce targeted changes into the genome which is based on the principle of homologous recombination.Combined with the somatic cell nuclear transfer technology,it has become an only efficient and pratical strategy to precisely modify the genomic fragments in domestic animals.Prion is the pathogen that caused infectious spongiform encephalopathy.Up to now,Prnp knock-out mice and sheep were successfully generated.However,no Prp knockout cows have ever been produced so far.Therefore,the goal of this study is to find an effective strategy for construction of promoter-less knock-out vector, and consequently to provide Prp knocked-out somatic cells for the attempts to clone Prp knocked-out cattle.更多还原