Survivin靶向的短发夹RNA表达载体的构建

Construction of the shRNA expression vectors targeting survivin

目的:构建存活素(Survivin)基因短发夹RNA(short hairpin RNA,shRNA)重组质粒并测序鉴定,为下一步探索乳腺癌基因治疗的RNA干扰途径建立基础。方法:设计并合成有小发夹结构的8条DNA片段,退火成互补双链后克隆至pSilencer adeno 1.0-CMV载体中构建重组质粒,转化大肠杆菌DH5α菌株,提取质粒行酶切鉴定后测序分析。结果:酶切鉴定和测序结果证实构建成功。结论:survivinshRNA重组质粒的成功构建为下一步用腺病毒包装,并感染荷瘤裸鼠行靶向RNA干扰抗乳腺癌的研究打下基础。

Objective：To construct the short hairpin RNA（shRNA） expression vector targeting survivin then identify it, which will make the foundation of exploring RNAi-based gene therapy for breast cancer. Methods： Eight strands of DNA fragement with the structure of survivin shRNA were designed and synthesized. After annealed, DNA strands were cloned into the pSilencer adeno 1. 0-CMV system, then transformed DH5a with the recombinants. The plasmids were amplified and extracted so as to identify by electropberesis and sequencement. Results： We found the expression vectors were constructed successfully. Conclusion： The construction of the vector paved a way for the further research, in which the recombinants will be packed with adenovirus and infect nude mice bearing human breast cancer.