针对livin的siRNA表达载体的构建

Construction livin-specific siRNA expression vector

目的设计能够有效抑制livin基因表达的小干扰RNA,构建针对livin基因的siRNA重组表达载体。方法通过分子克隆技术,设计并合成具有基因特异性的一组寡核苷酸片段,克隆到pSilencerTM3.1-H1 hygro载体,并经BamHⅠ/EcoRⅠ双酶切及测序鉴定证实,livin的siRNA序列成功地克隆到表达载体中。结果经BamHⅠ/EcoRⅠ双酶切及测序鉴定证实,人livin的siRNA序列成功地克隆到表达载体pSilencerTM3.1-H1 hygro中,构建了表达载体,测序分析证实插入序列正确。结论成功构建了livin表达载体,为后续研究奠定基础。

Objective The study aims to design a siRNA which could inhibit livin gene and construct eukaryotic ex- pression vector. Methods A group of double strand oligonucleotide fragments were synthesized and cloned into psi- leneerTM 3. l-H1 hygro vector with molecular cloning technique. It was confirmed by restrictive enzymes （ BamH I/EcoR I ） digestion and analysis of DNA sequencing. Results Restrictive enzymes digestion analysis and DNA se- quencing encoding livin was exactly the same as the reported sequence of encoding human livin in GenBank. The re- combinant plasmid was constructed. Conclusion The expression vector of livin were constructed successfully which lay the foundation for next study.