title: [Construction and identification of adenovirus vector for double-mutant human hypoxia-inducible factor-1alpha gene].
abstract: 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.
authors.affiliation: Department of Cardiology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. tongkai0521@sohu.com
authors.forename: Kai
authors.initials: K
authors.lastname: Tong
authors.forename: Yi-Jun
authors.initials: YJ
authors.lastname: Xie
authors.forename: Yue-Gang
authors.initials: YG
authors.lastname: Wang
authors.forename: Shou-Gui
authors.initials: SG
authors.lastname: Guo
authors.forename: Wen-Yan
authors.initials: WY
authors.lastname: Lai
authors.forename: Ping-sheng
authors.initials: PS
authors.lastname: Wu
mesh_terms: 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
chemical_list: D051795:Hypoxia-Inducible Factor 1, alpha Subunit