重组前强啡肽基因35型腺病毒载体的构建

Construction of the recombinant adenovirns vector with fibers derived from B group serotype 35 （rAd5/F35） containing prodynorphin gene

目的构建含前强啡肽（PDP）基因的重组35型腺病毒载体（Ad5／F35）。方法以pUC57-PDP重组质粒为模板扩增PDP基因，将回收的聚合酶链反应（PCR）产物片段克隆入pDC316载体，获得重组质粒pDC316-PDP。骨架质粒pBHG—fiber5／35和穿梭质粒pDC316-PDP共转染293细胞，同源重组产生Ad5／F35-PDP。经PCR鉴定目的基因的表达。结果PCR表明Ad5／F35-PDP质粒构建正确。结论获得的Ad5／F35-PDP可以用于转基因治疗的实验研究。

Objective To construct the recombinant adenovirus vector with containing fibers derived from B group serotype 35 （ rAdS/F35 ） containing prodynorphin gene. Methods The PDP gene was amplified with the pUC57-PDP plasmid as a formwork. After the purification, the gene fragment was cloned into the pDC316 carrier for the recombination of the plasmid of pDC316-PDP. The 293 cells were co-transfected by the skeleton plasmid of pBHG-fiberS/35 and the shuttle plasmid of pDC316-PDP, and the recombinant plasmid of AdS/F35-PDP was obtained. The expression of the transfected genes was detected by PCR and immunocytochemistry. Results The identification of PCR and immunocytochemical stain showed that the construction of the recombinant Ad5/F35-PDP plasmid could be confirmed. Conclusion The rAdS/F35-PDP vector can be used in empmcal study of transgenie therapy.