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


CONCLUSIONS
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.
Tong
Kai
K
Department of Cardiology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. tongkai0521@sohu.com
Xie
Yi-Jun
YJ
Wang
Yue-Gang
YG
Guo
Shou-Gui
SG
Lai
Wen-Yan
WY
Wu
Ping-sheng
PS
D000256:Adenoviridae / Q000235:genetics*; D045744:Cell Line, Tumor; D003327:Coronary Disease / Q000628:therapy; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000096:biosynthesis*; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics*; D010957:Plasmids
D051795:Hypoxia-Inducible Factor 1, alpha Subunit