Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (7): 1169-1173.doi: 10.3969/j.issn.1673-8225.2011.07.007

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Expression of recombinant human interleukin-1 receptor antagonist protein fluorescent plasmid in chondrocytes

Zhang Ping1, Liu Bin2, Cai Dao-zhang2, Zhong Zhi-hong1, Pan Yong-qian1, Zhang Zhen-shan1   

  1. 1Department of Orthopaedics, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou  510150, Guangdong Province, China
    2Department of Orthopaedics, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou  510630, Guangdong Province, China
  • Received:2010-09-20 Revised:2010-11-05 Online:2011-02-12 Published:2011-02-12
  • About author:Zhang Ping☆, Doctor, Associate chief physician, Department of Orthopaedics, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, Guangdong Province, China 644459349@qq.com
  • Supported by:

    the Scientific and Technological Project of Guangdong Province, No.  (2009)198*, the Doctor Fund of Guangzhou Medical University, No.2008C20*

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Abstract:

BACKGROUND: Interleukin-1 receptor antagonist (IL-1Ra) can delay osteoarthritis progress. The expression of IL-1Ra can be increased by gene transfection.
OBJECTIVE: To construct recombinant human IL-1Ra protein fluorescent plasmid and to observe its expression in chondrocytes by liposome gene transfection. 
METHODS: pGEM-T-IL-1Ra was digested with double enzymes restriction, and then it was linked with T4 DNA ligase and cloned into the pEGFP-C1 vector. Rabbit articular chondrocytes were isolated and cultured in vitro. The plasmids carrying the IL-1Ra gene were transfected into chondrocytes. Then the expression of the transgene was observed under a fluorescence microscope and detected by using real-time PCR assay.
RESULTS AND CONCLUSION: The sequence of obtained pEGFP-C1-IL-1Ra was identical to IL-1Ra and sequence in the Genbank. The expression of enhanced green fluorescent protein was observed. Real-time PCR analysis showed that the genes were expressed in chondrocytes.

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