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《Sichuan Journal of Physiological Sciences》 2008-03
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Construction of the eukaryotic expression plasmid for the cytokine receptor-chimeric antibody

Wu Jing1,Zhang Ping1,Zhou Chang-hua2,Zhao Zong-rong1,Wei Da-peng1,Zhang Chong-jie1 (1.Department of Immunology,College of Preclinical and Forensic Medicine,Sichuan University;2.Chengdu Blood Center,Chengdu 610041,China)  
Objective: To construct cytokine receptor-anti human AFP chimeric antibody gene.Methods:Extracellular region of EpoR gene and transmembrane domain of gp130 gene were cloned by RT-PCR method from hepatic tissue of fetus,and the two genes were spliced by endonuclease digestion.VL gene was cloned from anti-human AFP ScFv gene,and was spliced with Human CК gene to construct chimeric light chain.VH gene was cloned from anti-human AFP ScFv gene,and ligated into pAH4604 containing human γ1 gene to construct VH-CH gene.EpoR-gp130 gene and VH-CH gene were spliced by endonuclease digestion,and cloned into pVITRO-L to construct pVITRO-E-g-Ig.Results:Extracellular region of EpoR gene is 700bp,transmembrane domain of gp130 gene is 900bp.The recombinant gene was confirmed to be correct by restriction enzyme digestion and DNA sequencing.Conclusion: The cytokine receptor-anti human AFP chimeric antibody fusion protein was constructed and expressed successfully.
【Fund】: 四川大学创新基金(2004CF09)
【CateGory Index】: R346
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