PML重组腺病毒载体的构建与鉴定

CONSTRUCTION AND IDENTIFICATION OF RECOMBINANT ADENOVIRUS CONTAINING HUMANPML GENE

目的:构建携带PML目的基因的重组腺病毒载体(Ad-pml)。方法:应用基因重组技术将PSG-CMV和PSG5-PML分别用EcoRI和BglII双酶切,构建PSG-PML载体。将质粒PSG-PML与含有5型腺病毒右臂的质粒pPE3共转染至293细胞,产生含PML基因的重组腺病毒(Ad-pml),并经PCR及DNA测序鉴定。结果:经PCR反应可扩增出1767bp大小的片段,测序结果检测Ad-pml序列正确,通过免疫荧光反应证明病毒包装成功并且具有感染力。结论:成功构建了重组腺病毒载体(Ad-pml),为研究肿瘤基因治疗提供了基础。

Objective：To construct the recombinant adenovirus plasmid carring human PML gene for further study. Methods：By gene recombinantion technique, the plasmid of PSG- CMV and PSG5 -PML, double -digested with restrictive endonucleases EcoRI and BglII, constructed PSG - PML vector. Finally, the PSG - PML recombiiaant plasmid was transfected into 293 cell with pPE3 containing Adenovirus type 5 that including Right Arm , which produced Ad - pml. Ad - pml was verified by PCR and DNA sequence. Results： Identification of the Ad - pml by restriction digestion and PCR showed the length. Ad - pml was confirmed in sequence, fluorescence showed successful recombined adenovirus with infectiveness. Conclusion ： Ad - pml was successfully constructed. The development of Ad - pml will provide the basis for the research and therapy of cancer.