乙肝病毒PreS2基因在大肠埃希菌中的表达  

Expression of PreS2 gene from hepatitis B virus in Escherichia coli

作　　者：邵丽军[1] 李猛[2] 张利芳[3] 刘照惠[4] 

机构地区：[1]潍坊医学院预防医学系,山东潍坊261042 [2]潍坊医学院临床学院,山东潍坊261042 [3]包头医学院医学技术系,内蒙古包头014010 [4]长春生物制品研究所疫苗室,吉林长春130062

出　　处：《中国病原生物学杂志》2008年第6期417-420,共4页Journal of Pathogen Biology

摘　　要：目的在大肠埃希菌(Escherichiacoli)中表达乙肝病毒前S2(HBVPreS2)蛋白,并对其进行鉴定和纯化。方法利用DNA重组技术,将全长的HBVPreS2基因克隆至pMAL-C2x[融合表达载体含麦芽糖结合蛋白(maltosebindingprotein,MBP)标签蛋白]质粒中,构建pMAL-C2/S2表达载体,转化至E.coli,经IPTG诱导表达,SDS-PAGE分析目的蛋白的表达形式,依据其不同的表达形式采取适当的纯化方案,纯化产物经Westernblot和ELISA法检测反应原性,经Xa因子切割去除MBP标签蛋白。结果成功构建了表达载体pMAL-C2/S2,目的蛋白为PreS2-MBP,以包涵体和可溶性2种形式表达。纯化产物能与anti-HepatitisBvirusPreS2Antigen发生特异性反应。融合蛋白经Xa因子切割去除了MBP标签蛋白。结论在E.coli中成功表达了HBVPreS2蛋白,为进一步研制新型HBV预防及免疫治疗制剂打下了基础。Objective To express hepatitis B virus （HBV） PreS2 fusion protein in Escherichia coli. The expression product was identified and purified. Methods The total HBV PreS2 protein gene was inserted into prokaryotic expression vector pMAL-C2x by DNA recombination technique. The recombinant plasmid was named pMAL-C2/S2 fusion vector. pMAL-C2/S2 was induced by IPTG in E. coli. The expression products were analyzed by SDS-PAGE. The fusion protein was purified by anion-exchange chromatography and Amylose Resin affinity chromatography. The furified protein was identified by Western blot and ELISA. The MBP tap protein of fusion protein was removed by Xa factor. Results The recombinant prokaryotic expression vector pMAL-C2/S2 was successfully constructed. The interest protein was HBV PreS2-MBP fusion protein and were expressed as inclusion body and soluble form. Western blot and ELISA results showed that fusion protein could react with anti-hepatitis B virus PreS2 antigen. The MBP tap protein of fusion protein was removed by Xa factor. Conclusion HBV PreS2 protein has been successfully expressed in E. coli and the resulted protein can be used in future as neotype preventive or immunotheraDv reagent.

关 键 词：大肠埃希菌 表达 乙型肝炎病毒 PRES2蛋白 

分 类 号：R378.21[医药卫生—病原生物学] R373.21[医药卫生—基础医学]