单纯疱疹病毒II型糖蛋白G主要抗原表位的表达和抗原性检测  

Expression and Antigenic Analysis of the Recombinant Epitope of Herpes Simplex Virus Type 2 Glycoprotein G

作　　者：王晓红[1] 卢海蓉[1] 章刚[1] 陈少娟[1] 黄德新[1] 李凌云[1] 林枫[1] 

机构地区：[1]深圳大学生化工程技术研究中心,深圳518057

出　　处：《中国生物工程杂志》2007年第9期63-68,共6页China Biotechnology

摘　　要：利用PCR拼接技术，合成含单纯疱疹病毒II型糖蛋白G（gG2）抗原表位（氨基酸序列第561～578位）的片段，并进一步利用基因工程技术获得该表位的双拷贝片段，克隆入pET—KDO表达载体进行原核表达。经IPTG诱导后，高效表达出分子量大小约为39kDa的融合蛋白，经Western blot检测具有良好的抗原性。表达的融合蛋白经凝血酶切割和亲和层析纯化，得到双拷贝gG2（561～578aa）目的蛋白，经ELISA检测具有良好的灵敏度和特异性。该重组抗原的构建和表达可用于HSV-2特异性血清学诊断的研究。A fragment containing amino acid residues 561 - 578 of HSV-2 glycoprotein G （gG2） was obtained by PCR assembling technique, and doubly cloned into vector pET-KDO. The recombinant plasmid was transformed to BL21 （ DE3 ） plysS. Fusion protein, of molecular weight about 39kDa was highly expressed by induction of IPTG. Western blot result showed the fusion protein had good antigenicity. After putification and digestion,the purity reached 95%. The digested purified protein was analysed by ELISA and showed good sensitivity and specificity. The recombinant protein should be useful for type-specific serodiagnosis of HSV-2.

关 键 词：人单纯疱疹病毒 糖蛋白G 抗原表位 

分 类 号：Q819[生物学—生物工程]