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《Acta Academiae Medicinae Militaris Tertiae》 2006-13
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Identification and characterization of a novel hepatitis B virus core antigen binding protein

LU Yin-ying,YANG Yong-ping,WANG Lin,LI Ke,LIU Yan,ZHANG Ling-Xia,CHENG Jun(Gene Therapy Research Center,Institute of Infectious Diseases,The 302 Hospital of PLA,Beijing 100039,China)  
Objective To elucidate the biological function of hepatitis B virus core antigen(HBcAg) on pathogenesis of hepatitis B and to identify and characterize the gene coding for a novel protein interacting with HBcAg in hepatocytes.Methods The yeast two-hybrid screening was performed to isolate cDNA clones encoding a novel proteins(C12) that directly interact with HBcAg.The interaction between HBcAg and C12 was verified by re-mating.The fluorescence protein fusion gene of pEGFP-N1-C12 was transfected into 293 cells and L02 cells and observed by fluorescent microscope.MTT assay was used to study the effect of C12 on metabolism of mammal cells.Yeast two-hybrid and cDNA microarray were performed to search binding protein and differential expression gene adjusted by C12.Results C12 gene was screened and identified by yeast two-hybrid system 3.The interaction between HBcAg and the novel protein coded by the new gene C12 was further confirmed by re-mating.After 48 h,fluorescence could be observed steadily in cell plasma.MTT assay showed that the C12 expression did not influence the growth of liver cells.Seventeen differential expression genes in HepG2 cells transfected with C12 protein expression plasmid by cDNA microarray were screened out,of which 16 genes were upregulated and 1 gene were downregulated.Twenty-one colonies containing 16 different genes coding for C12 binding proteins were isolated by yeast two-hybrid,2 new genes with unknown function.Conclusion The novel protein C12 locates in cell plasma and interacts with many proteins in hepatocytes and may involve in many process of gene expression.C12 overexpression has no significant effect on the metabolism of liver cell.
【CateGory Index】: R392
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