携带HIV-1vpr基因的重组腺病毒的构建与鉴定

目的　构建携带HIV 1vpr基因的重组腺病毒 ,为进一步基因治疗的实验研究奠定基础。方法　利用AdEasy 1系统 ,通过含有目的基因片段vpr的穿梭载体pShuttle CMV/vpr和骨架质粒pAdEasy 1在细菌内同源重组的方法构建重组腺病毒质粒pAdCMV/vpr。将重组质粒转染 2 93细胞包装出重组腺病毒rvAdCMV/vpr。利用PCR等方法对重组腺病毒进行鉴定。结果　获得表达vpr蛋白的重组腺病毒。结论　成功构建出携带HIV 1vpr基因的重组腺病毒 ,为更深入的基因治疗研究奠定了基础。

Construction and Identification of Recombinent Adenovirus Carrying Vpr Gene

Objective To construct a recombinant adenoviral vector carrying HIV 1 vpr gene for the gene therapy research of malignant tumors.Methods The AdEasy adenoviral vector system was used in this experiment. Vpr gene was linked to pShuttle CMV plasmid and then homologous recombination was performed in E. coli BJ5183 between pShuttle CMV/vpr and adenovirus backbone plasmid pAdEasy 1. The obtaining recombinant vector was transfected into 293 cells and packaged out recombinant adenovirus rvAdCMV/vpr.Results The results of identifyication showed the insertion was right and the recombinant adenovirus contained the vpr gene. Conclusion The successful construction and the method for recombination lay an important foundation for the further study.