中国药理学通报2011，Vol.27Issue(4)：462-467,6.DOI:10.3969/j.issn.1001-1978.2011.04.006
利用Gateway技术构建重组腺病毒pAd-NK4
Gateway technical supported construction of a recombinant adenovirus vector pAd-NK4
摘要
Abstract
Aim To construct a recombinant adenovirus vector pAd-NK4 with Gateway technology. MethodsThe NK4 gene was amplified by PCR with plasmid containing hepatocyte growth factor ( HGF ) / NK 4 as ternplate, and then cloned into adenovirus adshuttle vector pYr-adshuttle-6 to acquire recombinant plasmid pYr-adshuttle-6-NK4, from which, the expression cassette of NK4-IRES-EGPF was transfered to pAD/BL-DEST with LR recombinant reaction to acquire recombinant adenovirus plasmid pAd-NK4-IRES-EGFP. The correct clone was linearized and transfected into 293 A cells for packing and amplified adenovirus pAd-NK4 was obtained and identified. The titer was determined by TCID50. The transfection efficiency was observed under the fluorescent microscope. Results The recombinant adenovirus vector containing human NK 4 gene was constructed successfully with the titer 6. 3 × 10 PFU · L . This vector was found to infect HEK293 cells efficiently. Conclusion The recombinant adenovirus expressing NK4 gene and green fluorescence can be successfully constructed with Gateway technology , which lays the foundation for further study.关键词
腺病毒/Gateway技术/人肝细胞生长因子/NK4/基因治疗/肿瘤分类
医药卫生引用本文复制引用
阙文忠,陈君敏..利用Gateway技术构建重组腺病毒pAd-NK4[J].中国药理学通报,2011,27(4):462-467,6.基金项目
国家自然科学基金资助项目(No 30871111) （No 30871111）
