期刊文献+

Construction of adeno-associated virus coexpression system for human angiopoietin-1 and VEGF gene 被引量:3

Construction of adeno-associated virus coexpression system for human angiopoietin-1 and VEGF gene
原文传递
导出
摘要 Background Ischemic disease is one of the leading causes of death in the world. In order to further study gene therapy for ischemic disease, we constructed a recombinant plasmid for co-expression of human angiopoietin-1 and vascular endothelial growth factor 165(VEGF165) gene in adeno-associated virus (AAV) gene delivery system. Methods Human angiopoietin 1 and VEGF165 gene were obtained using PCR. The upstream of angiopoietin 1 contained restriction enzyme site HindⅢ, and the downstream of angiopoietin 1 contained restriction enzyme site BamHⅠ. The upstream of VEGF165 contained restriction enzyme site BglⅡ, and the downstream of VEGF165 contained restriction enzyme site BamHⅠ. Using the multiple cloning sites (MCS) in plasmid pZero++ such as BamHⅠ, BglⅡ, HindⅢ, NotⅠ, XhoⅠ, XbaⅠ, SalⅠ, BspHⅠ, KspⅠ and the corresponding MCS in plasmid pAAV-MCS, angiopoietin 1 and VEGF165 gene were subcloned into pAAV-MCS. Results DNA sequencing revealed that the PCR- amplified angiopoietin 1 and VEGF165 were consistent with NCBI Gene Bank. The recombinant plasmid was identified using PCR and digestion, which proved to be consistent with our hypothesis. In recombinant plasmid, angiopoietin1 and VEGF possessed a CMV promoter and polyA terminator system respectively, thus assuring co-expression of the two genes. Conclusion Successful construction of AAV co-expression system for human angiopoietin 1 and VEGF165 gene will provide the foundation for gene therapy to cure severe ischemic disease. Background Ischemic disease is one of the leading causes of death in the world. In order to further study gene therapy for ischemic disease, we constructed a recombinant plasmid for co-expression of human angiopoietin-1 and vascular endothelial growth factor 165(VEGF165) gene in adeno-associated virus (AAV) gene delivery system. Methods Human angiopoietin 1 and VEGF165 gene were obtained using PCR. The upstream of angiopoietin 1 contained restriction enzyme site HindⅢ, and the downstream of angiopoietin 1 contained restriction enzyme site BamHⅠ. The upstream of VEGF165 contained restriction enzyme site BglⅡ, and the downstream of VEGF165 contained restriction enzyme site BamHⅠ. Using the multiple cloning sites (MCS) in plasmid pZero++ such as BamHⅠ, BglⅡ, HindⅢ, NotⅠ, XhoⅠ, XbaⅠ, SalⅠ, BspHⅠ, KspⅠ and the corresponding MCS in plasmid pAAV-MCS, angiopoietin 1 and VEGF165 gene were subcloned into pAAV-MCS. Results DNA sequencing revealed that the PCR- amplified angiopoietin 1 and VEGF165 were consistent with NCBI Gene Bank. The recombinant plasmid was identified using PCR and digestion, which proved to be consistent with our hypothesis. In recombinant plasmid, angiopoietin1 and VEGF possessed a CMV promoter and polyA terminator system respectively, thus assuring co-expression of the two genes. Conclusion Successful construction of AAV co-expression system for human angiopoietin 1 and VEGF165 gene will provide the foundation for gene therapy to cure severe ischemic disease.
出处 《Chinese Medical Journal》 SCIE CAS CSCD 2004年第4期562-565,共4页 中华医学杂志(英文版)
基金 TheprojectwassupportedbyNationalNatureScienceFoundationofChina (No 3 0 2 712 65 )
关键词 ANGIOPOIETIN-1 VEGF (165) adeno-associated virus plasmids angiopoietin-1 · VEGF (165) · adeno-associated virus · plasmids
  • 相关文献

参考文献11

  • 1Joseph R. Smith-Arica PhD,Jeffrey S. Bartlett PhD.Gene therapy: Recombinant adeno-associated virus vectors[J].Current Cardiology Reports.2001(1)
  • 2ClaussM.FunctionsoftheVEGFreceptor1 (FLT 1)inthevasculature[].TrendsCardiovascMed.1998
  • 3AsaharaT,ChenD,TakahashiT etal.Tie2receptorligands angiopoietin1andangiopoietin2,modulateVEGF reducedpostnatalneovascularization[].Circulation Research.1998
  • 4ZhangZ,DongH,LiuJ etal.Vascularendothelialgrowthfactorgenetransferimproveshostendothelializationofxenogeneicbiologicheartvalveinvivo[].Chinese Medical Journal.2002
  • 5BaumgartnerI,PieczekAR,ManorO etal.ConstitutiveexpressionofphVEGF165afterintramusculargenetransferpromotescollateralvesseldevelopmentinpatientswithcriticallimbischemia[].Circulation.1998
  • 6Ning Cong et al Dept pediatr tongji hospital tongji meduni Wuhan 430030.PEDIATRICS[J].Chinese Medical Journal,1995(11):65-65. 被引量:3
  • 7Risau W: Mechanisms of angiogenesis. Nature . 1997
  • 8Sundberg C,Kowanetz M,Brown LF,et al.Stable expression of angiopoietin-1 and other markers by cultured pericytes: phenotypic similarities to a subpopulation of cells in maturing vessels during later stages of angiogenesis in vivo[].Laboratory Investigation.2002
  • 9Ferrara N.Davis-Smyth T: The biology of vascular endothelial growth factor[].Endocrine Reviews.1997
  • 10Valenzuela DM,Griffiths JA,Rojas J,et al.Angiopoietins 3 and 4: diverging gene counterparts in mice and humans[].Proceedings of the National Academy of Sciences of the United States of America.1999

共引文献2

同被引文献5

引证文献3

二级引证文献8

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部