乙型肝炎病毒前S1蛋白反式激活蛋白2结合蛋白1基因的克隆及其反式调节基因的筛选  

Identification of genes regulated by HBV preSl-transactivated protein 2 binding protein 1

作　　者：张桓虎[1] 王丹琼[1] 成军[2] 赵婕[1] 赵心恺[1] 高虹[1] 

机构地区：[1]山西医科大学第二医院肝病中心,太原030001 [2]北京地坛医院

出　　处：《中华肝脏病杂志》2010年第4期267-270,共4页Chinese Journal of Hepatology

摘　　要：目的克隆HBV前S1蛋白反式激活蛋白2结合蛋白I（PS1TP2BP1）基因，并筛选与其相互作用的反式激活基因。方法应用聚合酶链反应（PCR）扩增PS1TP2BP1基因，鉴定正确后再将其亚克隆到真核表达载体pcDNA^TM 3．1／myc-His A上；以真核表达质粒pcDNA^TM 3．1／myc HisA—PS1TP2BP1转染HepG2细胞，构建cDNA消减文库；进行测序及同源性分析。结果从HepG2细胞来源的cDNA中扩增出PS1TP2BP1基因，并成功进行TA克隆，酶切、测序均正确后，成功构建真核表达重组体。消减文库扩增后得到35个阳性克隆，经菌落PCR分析，得到15个含200～1000bp插入片段的菌落。对所得片段测序，并进行同源性分析，显示15种已知基因编码蛋白可能是PS1TP2BP1反式激活靶基因。结论成功克隆PS1TP2BP1，并构建了PS1TP2BP1反式激活基因差异表达的cDNA消减文库，为今后进一步分析、研究病毒蛋白的致病机制奠定了基础。Objective To identify genes regulated by HBV preS 1-transactivated protein 2 binding protein 1 （PS1TP2BP1）. Methods PS1TP2BPI gene was amplified by polymerase chain reaction （PCR） technique and cloned into the eukaryotic expression vector pcDNA^TM 3.1/my-c-His A. The mRNAs isolated from HepG2 cells transfected recombinant eukaryotic expression vector pcDNATM3.1/myc- HisA-PS 1TP2BP 1 and pcDNATM3.1/myc-HisA empty vector were used to construct subtractive library. The differentially expressed genes were idenfied and analyzed. Results 35 differentially expressed clones were obtained. Colony PCR identified 15 clones with 200-1000 bp inserts. Sequence analysis identified 15 differentially expressed genes. Conclusion This study provides data for further characterize the function of PS 1TP2BPI.

关 键 词：肝炎病毒 乙型 克隆 分子 抑制性消减杂交 PS1TP2BP1 

分 类 号：R3[医药卫生—基础医学]