Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (50): 9428-943.doi: 10.3969/j.issn.1673-8225.2011.50.029

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Construction and expression of Ad-EGFP-MCP-1 vector

Zhang Fang1, Wang Wei2, Xie Yue2, Zhang Hao2   

  1. 1Department of Rheumatology and Clinical Immunology, Jiangsu Provincial Institute of Traditional Chinese Medicine, Nanjing   210028, Jiangsu Province, China
    2Department of Cardiothoracic Surgery, Affiliated Hospital of Xuzhou Medical College, Xuzhou   221002, Jiangsu Province, China
  • Received:2011-05-19 Revised:2011-08-26 Online:2011-12-10 Published:2011-12-10
  • About author:Zhang Fang★, Master, Associate professor, Department of Rheumatology and Clinical Immunology, Jiangsu Provincial Institute of Traditional Chinese Medicine, Nanjing 210028, Jiangsu Province, China zhangfang1doctor@126.com Wang Wei, Doctor, Associate professor, Department of Cardiothoracic Surgery, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China wangwei-doctor@163.com Zhang Fang and Wang Wei contributed equally to this study.
  • Supported by:

    the National Natural Science Foundation of China, No.30571845*; the Natural Science Foundation of Jiangsu Province, No. BK2007033*; Science and Technology Development Foundation of Xuzhou City, No. XM09B072*

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Abstract:

BACKGROUND: Monocyte chemotactic factor can promote arterialization of newly formed vessels to form functional arteriole and then realize angiogenesis.
OBJECTIVE: To construct Ad-EGFP-MCP-1 vector and to observe its expression in vivo after package, purification and viral titer detection.
METHODS: MCP-1 gene was amplified by PCR and the sequence was compared with Genebank data, and then sub-cloned into the pDC315-EGFP vector after sequence analysis. Ad-EGFP-MCP-1 was obtained with pBHG lox ΔE1,3 Cre secondary packaging system. After transfection of Ad-EGFP-MCP-1 via trachea, EGFP expression was observed under fluorescence microscope. 
RESULTS AND CONCLUSION: The full-length of MCP-1 was obtained by PCR and identified by sequencing. Ad-EGFP-MCP-1 recombinant adenovirus vector was stably expressed in rat lung tissue. EGFP expression reached the peak level at 1 week and maintained this level for approximately 4 weeks. Ad-EGFP-MCP-1 recombinant adenovirus vector was successfully constructed, packaged and amplified, and it is effectively expressed in the lung tissue.

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