Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (41): 7728-7732.doi: 10.3969/j.issn.2095-4344.2012.41.026

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Construction and identification of recombinant adenovirus vectors encoding human transforming growth factor-beta 1 gene

Zeng Zhao-xun, Yin Cheng-hui, Qiu Jun-qin, Chen Zong-xiong   

  1. Institute of Orthopedics, Fuzhou General Hospital of Nanjing Military Command, Fuzhou 350025, Fujian Province, China
  • Received:2012-01-05 Revised:2012-02-01 Online:2012-10-07 Published:2012-10-07
  • Contact: Yin Cheng-hui, M.D., Associate chief professor, Institute of Orthopedics, Fuzhou General Hospital of Nanjing Military Command, Fuzhou 350025, Fujian Province, China chenghuiyin@hotmail.com
  • About author:Zeng Zhao-xun★, Studying for master’s degree, Institute of Orthopedics, Fuzhou General Hospital of Nanjing Military Command, Fuzhou 350025, Fujian Province, China

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Abstract:

BACKGROUND: Transforming growth factor-beta1 (TGF-β1) can promote the growth and proliferation of many cells and is one of main growth factors that regulate osteoblast differentiation of bone marrow mesenchymal stem cells.
OBJECTIVE: To construct a recombinant adenoviral vector carrying the human TGF-β1 gene using the AdMax system which may be applicable in gene therapy of bone defect.
METHODS: The hTGF-β1 gene sequence was amplified by PCR from cDNA template and then was inserted into shuttle plasmid pAV-MCMV-EGFP-3FLAG. The recombinant shuttle plasmid was constructed and transformed into E.coli DH5α.The recombinant pAV-MCMV-hTGF-β1 was obtained.
RESULTS AND CONCLUSION: The recombinant pAV-MCMV-hTGF-β1 was packaged and amplified in 293 cells. Then the recombinant TGF-β1 adenovirus was constructed successfully. After measuring the titre of virus (1.25×1010 pfu/mL), the target gene was evaluated by PCR and Western Blot. These findings suggest that construction of hTGF-β1 adenoviral vector was successfully achieved using the AdMax system and may be available in the future study of TGF-β1 gene therapy.

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