人巨细胞病毒PP150蛋白C端多肽与gp52蛋白片段的融合表达  

Expression of PP150 C-terminus and gp52 chimeric protein of human cytomegalovirus

作　　者：李越希[1] 陶开华[1] 汪兴太[2] 李长贵[2] 周宗安[1] 郁兴明[1] 黄培堂[3] 

机构地区：[1]南京军区军事医学研究所 [2]中国药品生物制品检定所 [3]军事医学科学院生物工程研究所

出　　处：《中华微生物学和免疫学杂志》2001年第5期556-559,共4页Chinese Journal of Microbiology and Immunology

摘　　要：目的　将人巨细胞病毒PP1 50蛋白C端多肽与gp52蛋白片段融合 ,构建表达该融合蛋白的工程菌。方法　合成PP1 50蛋白C端 2 5个氨基酸的基因片段 ,并选用细菌偏爱的密码子。用PCR方法扩增gp52蛋白基因片段。将这 2个基因片段分别克隆至同一质粒pET2 8a( + )内的NcoⅠ /BamHⅠ和BamHⅠ /EcoRⅠ位点 ,使两者串联在一起 ,并且翻译框架一致 ,可表达 1个融合蛋白。将重组质粒转化大肠杆菌 ,用酶切及SDS PAGE等方法筛选表达融合蛋白的工程菌。结果　构建成功了高效表达PP1 50C端多肽与gp52蛋白片段的融合蛋白工程菌 ,表达量为 3 5%左右。初步纯化获得了表达的融合蛋白。结论　初步纯化的融合蛋白能与已知的抗gp52 IgM阳性血清和抗PP1 50 IgM阳性血清反应 ,说明融合蛋白C端的gp52蛋白保持较好的抗原性 ,融合蛋白N端的PP1 50C端多肽也显示有抗原性。Objective To make up an engineering E.coli strain for expression of protein PP150 C terminus and gp52 protein of human cytomegalovirus. Methods The gene fragment of protein PP150 C terminal 25 amino acid was synthesized and cloned into NcoⅠ/BamHⅠ sites before gp52 gene fragment in plasmid pET28a+gp52, the recombinant plasmid was transformed into E.coli BL21(DE3) for screening the engineering E.coli expressing chimeric protein of PP150 and gp52, the recombinant plasmid was identified by restriction enzyme digestion, and the engineering E.coli was screened by SDS PAGE. The expressed chimeric protein was preliminarily purified and characterized by ELISA test. Results The engineering E.coli expressing PP150 and gp52 chimeric protein was established, and the chimeric protein has good antigenicity. Conclusion The expressed chimeric protein is potential for the detection of anti HCMV IgM, and is of great significance in the prevention of HCMV infection.

关 键 词：人巨细胞病毒 PP150蛋白 gp52蛋白 融合蛋白 基因表达 

分 类 号：R346[医药卫生—基础医学]