Livin基因siRNA重组表达载体的构建

Construction Livin-specific siRNA expression vector

目的构建针对人抑凋亡基因Livin的小干扰RNA(siRNA)重组表达质粒载体。方法通过分子克隆技术,设计并合成具有基因特异性的3组寡核苷酸片段,克隆到pmU6质粒载体,并经菌落PCR鉴定及测序鉴定,Livin的siRNA序列成功克隆到重组表达质粒载体中。结果经菌落PCR鉴定及测序鉴定证实,人Livin的siRNA序列成功克隆到表达载体中,插入序列正确。结论成功构建了Livin siRNA质粒表达载体,为后续研究降低或沉默Livin基因表达后能否有效抑制膀胱癌细胞增殖和促进细胞凋亡奠定了基础。

Objective The study aims to design and construct 3eukaryotic expression vectors of short hairpin RNA（shRNA）against Livin gene. Methods Three groups of double strand oligonucleotide fragments were synthesized and cloned respectively into pmU6 vector with molecular cloning technique.It was confirmed by colony PCR digestion and analysis of DNA sequencing. Results Colony PCR digestion and analysis of DNA sequencing,encoding Livin was exactly the same as the reported sequence of encoding human Livin in GenBank.The recombinant plasmid was constructed. Conclusions The expression vector of Livin-specific siRNA were constructed successfully which lay the foundation for subsequent study.