肿瘤相关基因hyrdC基因重组腺病毒的构建和鉴定

Experiment research of constructing and verifying recombinant adenovirus vector carrying hyrdC gene

目的利用AdEasy腺病毒载体系统构建携带hyrdC基因的高滴度重组腺病毒，为进一步研究hyrdC基因的功能及机制做准备。方法以逆转录聚合酶链反应（RT—PCR）法扩增hyrdC基因，构建腺病毒重组质粒pShutth—CMV—hyrdC，线性化后采用电穿孔法与腺病毒骨架质粒pAdEasy-1共同转化BJ5183感受态大肠杆菌，挑选重组质粒将其转染293细胞，获得高滴度重组腺病毒Ad—hyrdC。结果成功地制备了高滴度携带hyrdC基因的重组腺病毒。结论采用AdEasy腺病毒载体系统能够构建携带hyrdC基因的重组腺病毒，为进一步研究hyrdC基因的功能及机制奠定了基础。

Objective To construct recombinant adenovirus carrying human tumor related hyrdC gene by AdEasy adenovirus vector system for further studying on the function and mechanism of hyrdC gene. Methods HyrdC gene was amplified by RT - PCR, recombinant shuttle plasmid pShuttle - CMV -hyrdC was constructed, then the linearized pShuttle - CMV - hyrdC was dectroporated into E. coli BJ5183 with adenovirus backbone plasmid pAdEasy - 1. Selected recombinants were tnmsfected into HEK cells to acquire high titer recombinant adenovirus Ad - hyrdC. Results Recombinant adenovirus carrying hyrdC gene was constructed successfully, and the high titer recombinant adenovirns was acquired. Conclusion Recombinant adenovirns carrying hyrdC gene could be constructed and provided the basis for the further study of function and mechanism of hyrdC gene.