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Construction of Eukaryotic Expression Vector Containing Antiviral MxA Gene and Its Expression in Chicken Cells
Li Liu, Ke-Jun Cai, Yi-Xiang Zhang, Pai He1, Zhi-Li Ding, Nian-Ci Zhang*
School of Life Sciences, Huzhou Teachers College, Huzhou 313000, China; 1College of Life Sciences, Soochow University, Suzhou 215006, China
Abstract: Human MxA has broad antiviral spectrum. The aim of the study was to construct eukaryotic expression vector containing antiviral MxA gene which was expressed in chicken cells in order to study the role of MxA against avian virus disease. MxA gene was obtained from pCDNA3.1(+)-MxA by restriction enzyme EcoRI and ApaI， and linkaged with eukaryotic expression vector pEGFP-C1. The recombinant plasmid pEGFP-C1-MxA was identified by PCR and restriction analysis. pEGFP-C1-MxA was tranfected into chicken embryo fibroblast (CEF) and chicken testicle tissue cells. Granular green fluorescence was detected by fluorescence microscope in cytoplasm of transfected cells after 48 h， and specific fragments of MxA and EGFP gene were amplified respectively by RT-PCR with total RNA of transfected cells. Moreover， the expression of EGFP-MxA fusion protein was detected by immunohistochemistry with monoclonal antibody of GFP. The data indicated that the antiviral gene MxA of the recombinant plasmid could be expressed in CEF and chicken testicle tissue cells. The results would contribute to further studies of application of MxA in dealing with chicken virus diseases.
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