摘要
目的:构建并鉴定针对基因ATRN的特异性siRNA真核表达载体。方法:根据ATRN基因cDNA序列,设计针对目的基因ATRNcDNA序列的3个siRNA靶序列,将其插入H1启动子下游,克隆到真核表达载体psiRNA-hH1neo中,转化DH5α菌株扩增,提取质粒通过酶切鉴定和DNA测序鉴定。结果:酶切鉴定及DNA测序结果显示插入片断正确。结论:本研究成功构建针对ATRN的特异性真核表达载体,为进一步研究ATRN基因在雄性生殖系统的功能奠定基础。
Objective: To construct and identify the siRNA eukaryotic expression vector targeting gene ATRN. The siRNAs were designed according to the coding the cDNA sequence of ATRN gene, and cloned into the downstream of H1 promoter of psiRNA - hHlneo. Then, the vector was transformed into DH5α strain. The constructed recombinant was analyzed and identified by restriction endonuclease digestion and DNA sequencing. Results: The constructed psiRNA plasmid digested with restriction endonuclease Ase I was linearized. The sequencing result confirmed that the sequence of inserted fragment was correct. Conclusion: Eu- karyotic expression vector of siRNA targeting ATRN gene is successfully constructed, the results of study lay the foundation for further studying on the function of ATRN in the male reproductive system .
出处
《中国计划生育学杂志》
北大核心
2008年第6期351-354,共4页
Chinese Journal of Family Planning
基金
国家自然科学基金资助项目(30570679)