FAK短发夹RNA干扰重组体的构建与鉴定  

Cloning and sequence analyzing of the recombinant plasmid affecting gene FAK translation by RNA interfering

作　　者：郝嘉[1] 郭红[1] 吕伟[1] 肖颖彬[1] 

机构地区：[1]第三军医大学附属新桥医院全军心血管外科中心,重庆400037

出　　处：《四川医学》2006年第10期991-992,共2页Sichuan Medical Journal

基　　金：国家自然科学基金(批准号:30100063)

摘　　要：目的本研究利用RNA干扰技术,以粘附斑激酶(focal adhesion kinase,FAK)为靶基因,设计构建重组体,并对重组体进行鉴定,为下一步探索心肌纤维化的新途径奠定基础。方法设计小发夹结构四条DNA序列,经退火成互补两对双链,再克隆到载体PAVU6+27中构建重组体,转化入JM109菌株,提取质粒进行酶切鉴定、测序分析。结果酶切鉴定和测序分析均提示FAK靶向RNA干扰重组体的构建成功。结论FAK靶向RNA干扰重组体的成功构建,为下一步利用该重组体沉默心脏成纤维细胞中FAK基因的表达,探索心肌纤维化机制打下基础。Objective For the further searching new gene therapy method of the myocardial fibrosis, the recombinant plasmid affecting gene FAK translation by RNA interfering was cloned and the nucleic acid sequence was analysed. Methods Four DNA sequences containing small hairpin structure were designed and synthesized. The complement form was obtained by annealing and inserted into vector PAVU6 ＋ 27. The recombinant plasmid was transformed into JM109 strains. Finally the plasmid identified by restriction enzyme was used for sequence analysis. Results The recombinant was cloned and the target sequence was obtained. Conclusion Successful cloning the recombinant make it possible searching new gene therapy method of the myocardial fibrosis.

关 键 词：FAK RNA干扰 重组体 心脏 

分 类 号：Q78[生物学—分子生物学]