Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (36): 6753-6757.doi: 10.3969/j.issn.2095-4344.2012.36.018

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Construction and identification of recombinant adenovirus vector containing CXC chemokine receptor 4 gene

Pan Ting-ming1, Xu Hao2, Chen Jian-mei2, Song Chen-yang3, Yao Xiao-dong2   

  1. 1Department of Orthopedics, Second People's Hospital of Fujian University of Traditional Chinese Medicine, Fuzhou 350003, Fujian Province, China;
    2Department of Orthopedics, Fuzhou General Hospital of Nanjing Military Region, Fuzhou 350025, Fujian Province, China;
    3Department of Orthopedics, Union Hospital, Fujian Medical University, Fuzhou 350001, Fujian Province, China
  • Received:2012-01-20 Revised:2012-02-28 Online:2012-09-02 Published:2012-09-02
  • Contact: Xu Hao, Chief physician, Professor, Master’s supervisor, Department of Orthopedics, Fuzhou General Hospital of Nanjing Military Region, Fuzhou 350025, Fujian Province, China
  • About author:Pan Ting-ming★, Master, Physician, Department of Orthopedics, Second People's Hospital of Fujian University of Traditional Chinese Medicine, Fuzhou 350003, Fujian Province, China 3467890@163.com

Abstract:

BACKGROUND: Recent studies have demonstrated that stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) is capable of promoting the migration of bone marrow mesenchymal stem cells (BMSCs).
OBJECTIVE: To construct recombinant adenovirus vector containing CXCR4 gene and provide the basis for further research of CXCR4 gene for application.
METHODS: The total RNA of mouse was extracted and CXCR4 gene which is 1080 bp in size was amplified by RT-PCR. Fragment containing CXCR4 was cloned into the shuttle vector pAdTrack-CMV that carried a green fluorescence protein (GFP) gene to generate a recombinant plasmid pAdTrack-CMV-CXCR4. The recombinant adenovirus pAdEasy-CMV-CXCR4 was transferred into HEK293 cells for packaging and amplification. The viral titer was determined and the insert of CXCR4 gene was verified by PCR method and western blot.
RESULTS AND CONCLUSION: The recombinant adenovirus vector (pAdEasy-CMV-CXCR4) was successfully constructed and the sequence was corrected by PCR and sequencing, which possesses high titers. Western blot showed CXCR4 protein expression in infected HEK293 cells was markedly higher than that in uninfected HEK293 cells.

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