Construction and identification of adenovirus vector for double-mutant human hypoxia-inducible factor-1alpha gene.
- Author:
Kai TONG
1
;
Yi-Jun XIE
;
Yue-Gang WANG
;
Shou-Gui GUO
;
Wen-Yan LAI
;
Ping-sheng WU
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; genetics; Cell Line, Tumor; Coronary Disease; therapy; Genetic Therapy; Genetic Vectors; biosynthesis; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; genetics; Plasmids
- From: Journal of Southern Medical University 2007;27(4):445-447
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct an adenovirus vector containing the double-mutant hypoxia-inducible factor-1alpha (HIF-1alpha) gene for exploring the therapeutic angiogenesis for coronary heart disease.
METHODSHuman 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 CONCLUSIONThe 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.
