Construction of human anti-HBc ScFv eukaryotic expression vector and expression of anti-HBc ScFv in HepG2 cells |
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| Authors: | TANG Zheng-hao ZANG Guo-qing MA Hui-hui YU Yong-sheng JIANG Hong LI Gang YAO Ji-lu |
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| Institution: | 1.Department of Infectious Diseases, Shanghai Jiaotong University Affiliated Sixth People's Hospital, Shanghai 200233, China;2.Department of Infectious Diseases, Sun Yat-sen University Affiliated Third Hospital, Guangzhou 510630, China |
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| Abstract: | AIM: To construct an eukaryotic expression vector of human single-chain variable fragment against hepatitis B virus core protein (anti-HBc ScFv) and detect its expression in HepG2 cells. METHODS: Anti-HBc ScFv genes were amplified from the plasmids abstracted from positive clone and inserted into pEGFP-c1 vector that contained green fluorescent protein gene. The recombinant plasmids were transfected into HepG2 cells, and resistant clones were obtained by G418 selection. The expression of the gene of fusion protein was determined by fluorescent invert microscope and ELISA. RESULTS: Recombinant plasmids were successfully constructed. The plasmid transfected HepG2 cells were obtained by G418 selection. Specific fluorescence was observed in HepG2 cells 48 hours after transfection. ELISA analysis confirmed the expression of anti-HBc ScFv in the cells. CONCLUSION: The construction of human anti-HBc ScFv eukaryotic expression vector and its expression in HepG2 cells lay the foundation for advanced research of intracellular anti-HBc ScFv. |
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