人血管内皮生长因子-D的重组腺病毒制备与鉴定

CONSTRUCTION AND CONFIRMATION OF HUMEN VEGF-D RECOMBINANT ADENOVIRUS

生物旁路是近年来缺血性心脏病治疗研究的一个新热点。本研究旨在构建人血管内皮生长因子 D重组腺病毒 ,为基因转染研究作准备。用重组粘粒和DNA TPC混合后 ,以磷酸钙共沉淀法转染 2 93细胞 ,获取重组腺病毒。重组粘粒插入方向及重组腺病毒均经酶切鉴定为正确。所得重组腺病毒为E1缺陷 ,可用于体内基因治疗研究。

Biological bypass”is a new hotspot in the reserch of CAD. VEGF-D recombinant adenovirus was constructed and confirmed before the studies of gene transfection. VEGF-D - pAxCAwt cosmid DNA and DNA-TPC were mixed and co-transfected to the 293 cell by calcium phosphate coprecipitation. The result of restriction showed that the direction of the insert in the cosmid is right and the recombinant adenovirus generated contains the gene of VEGF-D, and is E1 deleted and can be further used in vivo gene therapy.