Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (20): 3741-3744.doi: 10.3969/j.issn.1673-8225.2011.20.031

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Construction and identification of vascular endothelial growth factor 121 and bone morphogenetic protein-2 genes co-expressing recombinant adenovirus vector

Zhong Sheng1, Liu Dan-ping1, Liu Su-wei2, Li Xiao-yu, Li Chen1, Li Yuan4   

  1. 1Department of Bone and Joint Surgery, First Affiliated Hospital of Liaoning Medical University, Jinzhou   121000, Liaoning Province, China; 2Department of Anatomy, Liaoning Medical University, Jinzhou   121000, Liaoning Province, China; 3Second Department of Orthopaedics, West Hospital of Anshan City, Anshan   114012, Liaoning Province, China; 4Third Department of Neurology,First Affiliated Hospital of Liaoning Medical University, Jinzhou  121000, Liaoning Provine, China
  • Received:2010-11-28 Revised:2010-12-12 Online:2011-05-14 Published:2011-05-14
  • Contact: Liu Dan-ping, Doctor , Chief physician, Professor, Master’s supervisor, Department of Bone and Joint Surgery, First Affiliated Hospital of Liaoning Medical University, Jinzhou 121000, Liaoning Province, China liudanping2009@ sohu.com
  • About author:Zhong Sheng★, Studying for master’s degree, Department of Bone and Joint Surgery, First Affiliated Hospital of Liaoning Medical University, Jinzhou 121000, Liaoning Province, China ciguangqingran@yahoo.com.cn

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Abstract:

BACKGROUND: Vascular endothelial growth factor 121 (VEGF121) and bone morphogenetic protein-2 (BMP-2) play an important role in the development and formation of bones and vessels. At present, there are few reports about the treatment of pathogenesis of steroid-induced avascular necrosis of the femoral head (SANFH) by VEGF121 combined with BMP-2 gene in China.
OBJECTIVE: To construct VEGF121 and BMP-2 genes adenovirus shuttle plasmid pShuttle-CMV-V EGF121-IRES-BMP2.
METHODS: After Plasmid pShuttle-CMV-VEGF121-IRES-hrGFP-1 through Kpn I/Xba I, BMP-2 fragments were directionally connected to pShuttle-CMV-VEGF121-IRES. Simultaneous expression of two gene plasmid pShuttle-CMV-VEGF121-IRES was constructed, and injected with H5a cells expansion, planking, screening positive colonies, extracting plasmid, and then was undergo restriction analysis and sequencing. The correct adenovirus plasmid which has been constructed and confirmed after through BJ5183-AD-1 electroporation competent cells was undergo the electroporation, planking, screening positive colonies, extracting plasmid, and was undergo restriction analysis, PCR detection, and sequence analysis.
RESULTS AND CONCLUSION: Construction of the adenovirus shuttle plasmid was correct according to restriction analysis and nucleotide sequence detection confirmed. It is indicated that VEGF121 and BMP-2 genes co-expressing recombinant adenovirus vector can be constructed successfully in the experiment.

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