Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (24): 4505-4508.doi: 10.3969/j.issn.1673-8225.2012.24.028

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Construction and identification of a recombinant pcDNA4 His/Max human bone morphogenetic protein eukaryotic expression vector

Jiang Wei1, Hu Zhen-ming2, Hao Jie2, Gan Qiang1   

  1. 1Chongqing Medical University, Chongqing 400016, China;
    2Department of Orthopedics, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
  • Received:2011-10-20 Revised:2011-11-30 Online:2012-06-10 Published:2013-11-05
  • Contact: Hu Zhen-ming, Doctor, Professor, Department of Orthopedics, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China zhenminghu62@yahoo.com.cn
  • About author:Jiang Wei★, Studying for master’s degree, Chongqing Medical University, Chongqing 400016, China
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Abstract:

BACKGROUND: Adenovirus mediated method has been used numerously in human bone morphogenetic protein 9 (hBMP-9) induced osteogenesis researches, but the security is the most important problem. Eukaryotic expressing vector is a way to solve this question.
OBJECTIVE: To reconstruct a recombinant DNA pcDNA4 His/Max hBMP-9 eukaryotic expressing vector by amplifying hBMP9 gene and then cloning into pcDNA4 His/Max.
METHODS: Padtrack-cmv-hBMP-9 was amplified by PCR, and hBMP-9 gene was retrieved by electrophoresis. pcDNA4 His/Max was digested by NotⅠ, Kpn Ⅰand then the hBMP-9 gene was cloned into pcDNA4 His/Max. The recombinant plasmid was transformed by DH5α, clonal expansion, and purification, and then verified by sequencing.
RESULTS AND CONCLUSION: The cloned hBMP-9 gene was 1.3 kb long, having the same length and sequence of the gene that human possessed. Eukaryotic expressing vector of pcDNA4 His/Max hBMP-9 has been constructed successfully.

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