[Construction and identification of adenovirus vector for double-mutant human hypoxia-inducible factor-1alpha gene]

Kai Tong, Yi-Jun Xie, Yue-Gang Wang, Shou-Gui Guo, Wen-Yan Lai, Ping-sheng Wu
Nan Fang Yi Ke da Xue Xue Bao, Journal of Southern Medical University 2007, 27 (4): 445-7

OBJECTIVE: To construct an adenovirus vector containing the double-mutant hypoxia-inducible factor-1alpha (HIF-1alpha) gene for exploring the therapeutic angiogenesis for coronary heart disease.

METHODS: Human double-mutant HIF-1alpha cDNA was obtained from PCR of pShuttle-2-HIF-1alpha containing the mutant HIF-1alpha gene (564). The recombinant adenoviral plasmid containing mutant HIF-1alpha cDNA was constructed by ligation of recombinant pShuttle2 containing double-mutant HIF-1alpha cDNA and Adeno-X viral DNA, followed by its identification and transfection into adenoviral packaging cells HEK293 via lipofectamine 2000.

RESULT AND CONCLUSION: The recombinant pAdeno-HIF-1alpha was correctly constructed and verified by restriction endonuclease and DNA sequencing analyseis. This recombinant adenovirus containing the double-mutant HIF-1alpha gene may facilitate further investigation of mutant HIF-1alphagene therapy for coronary heart disease.

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