Prohibitin基因的shRNA真核表达载体的构建及鉴定
Construction and identification of shRNA eukaryotic expression vectors targeting to prohibitin gene
何谦 1张淑群 2赵丽华 1王春玲 1楚雍烈3
作者信息
- 1. 西安交通大学医学院,第二附属医院检验科,陕西,西安,710004
- 2. 西安交通大学医学院,第二附属医院肿瘤科,陕西,西安,710004
- 3. 西安交通大学医学院,免疫和微生物教研室,陕西,西安,710061
- 折叠
摘要
目的:构建靶向prohibitin基因的shRNA真核表达载体.方法:根据prohibitin mRNA序列,利用Ambion网上设计软件设计两条针对该基因不同靶点的shRNA干扰序列,克隆到线性化的质粒载体上,并进行酶切鉴定和测序.结果:酶切结果表明构建的shRNA已插入载体,经测序证明与设计一致.结论:成功构建两个靶向prohibitin基因的shRNA真核表达载体,为下一步研究prohibitin基因在肿瘤细胞中的作用和后续的的体内外实验奠定了基础.
Abstract
AIM: To construct 2 shRNA eukaryotic expression vectors targeting to prohibitin gene. METHODS: Finding the sequence of human prohibitin gene in Pubmed, the oligonucleotides of shRNA were designed by the software of Ambion Company then synthesized and directionally cloned into pGenesill. 1 vector. Then the recombinant vectors were extracted and confirmed by restriction enzyme and sequencing analysis. RESULTS: The restriction enzyme analyses demonstrated that shRNA had been inserted into vectors, and sequencing analyses demonstrated that their sequences were the same as the design. CONCLUSION: We have successfully constructed shRNA eukaryotic expression vectors targeting to prohibitin gene. It laid the foundations for the further study on the function of prohibitin gene in tumor cell and the future RNAi experimental research in vivo and in vitro.
关键词
prohibitin基因/RNA干扰/shRNA/真核表达载体Key words
prohibitin gene/RNA interference/shRNA/eukaryotic expression vector引用本文复制引用
出版年
2009
NETL
NSTL
维普
万方数据