单纯疱疹病毒1型糖蛋白B胞外区基因片段的克隆、表达及初步鉴定  

Cloning,Expression and Initial Identification of Extracellular Region Gene Fragment of Herpes Simplex Viruses l

作　　者：吕敏[1] 潘明洁[2] 杨华凤[3] 李越希[2] 

机构地区：[1]南京医科大学基础医学院生物化学与分子生物学系,江苏南京210029 [2]南京军区军事医学研究所,江苏南京210002 [3]南京医科大学公共卫生学院流行病与卫生统计学系,江苏南京210029

出　　处：《药物生物技术》2008年第2期90-93,共4页Pharmaceutical Biotechnology

基　　金：国家"863"计划项目资金资助(2006AA02A226)

摘　　要：构建单纯疱疹病毒1型糖蛋白B（HSV1gB）胞外区基因片段的重组原核表达质粒，并获得HSV1gB胞外区基因的表达。选用真核和原核细胞均偏爱的密码子，化学合成含信号肽序列的HSV1gB蛋白胞外区基因序列，用PCR方法扩增编码HSV1gB胞外区1～696aa的基因，利用DNA重组技术将其定向插入到原核表达载体pGEX4T-2上，转化大肠杆菌TG1菌株，经IPTG诱导表达及SDS-PAGE鉴定分析。SDS-PAGE检测显示，表达的HSV1gB／GST融合蛋白分子质量为96ku。利用亲和层析方法，纯化获得了表达的目的蛋白。ELISA结果表明表达产物具有较好的抗原性和特异性。可溶性重组HSV1gB蛋白的表达成功为单纯疱疹病毒亚单位疫苗的研究奠定了基础。To construct the recombinant plasmid expressing extracellular region gene fragment of herpes simplex viruses 1 glycoprotein B and to acquire the protein. The extracellular region of the herpes simplex viruses 1 glyco protein B gene was amplified by PCR and was cloned into plasmid pGEX4T-2 by T4 ligase. The ligated products were transformed into E. coli TG1. The transformants were induced by IPTG and identified by SDS-PAGE for screening the positive transformants. The recombinant plasmid HSVlgB-pGEX4T-2 expressing HSVlgB was successfully constructed, which could distinctively express the HSVlgB/GST. ELISA result showed that the expressed 96kd HSVlgB chimeric protein has good antigenicity and specificity, which have established the foundation for developing herpes simplex viruses subunit vaccine.

关 键 词：单纯疱疹病毒 糖蛋白B 克隆 表达 

分 类 号：Q7[生物学—分子生物学]