日本血吸虫毛蚴相关蛋白基因SjMP13的克隆表达及免疫学鉴定  

CLONING, EXPRESSION AND IMMUNOLOGICAL IDENTIFICATION OF TIlE 13 KDA MIRACIDIA ASSOCIATED PROTEIN OF SCHISTOSOMA JAPONICUM

作　　者：周艳萍[1] 吕志跃[1] 廖鸿兴[2] 李任远[2] 郑焕钦[1] 吴忠道[1] 

机构地区：[1]中山大学中山医学院寄生虫学教研室,国家教育部热带病防治重点实验室,广州510080 [2]中山大学中山医学院临床专业,广州510080

出　　处：《寄生虫与医学昆虫学报》2009年第2期65-70,共6页Acta Parasitologica et Medica Entomologica Sinica

基　　金：国家自然科学基金项目（No.30771888）;国家“863”项目（No.2006AA02Z444）;国家大学生创新性实验计划

摘　　要：利用日本血吸虫感染者唾液免疫筛选日本血吸虫虫卵eDNA文库，对获得的阳性克隆进行序列分析，其中一阳性克隆插入片段包含一个351bp的开放读码框，其编码116个氨基酸，预测分子量为12．9kDa，将其命名为sjMP13，该蛋白与血吸虫尾蚴相关蛋白（GenBank序列号：AF448823．1）同源性高达96％。将该序列克隆入原核表达质粒pET32a（＋）并转化宿主菌BIS21／DE3，经IPTG诱导表达后，其重组表达产物经亲和层析纯化，再用Western-blot进行免疫学分析，该序列的原核表达产物能被日本血吸虫感染者唾液以及血清识别，提示该分子具有诊断抗原的开发价值。Saliva of patients with schistosomiasis was sampled as the antibodies to screen Schistosoma japonicum egg eDNA library, and sequencing was performed with the positive clones obtained. One of the positive clones had an open reading frame which contained 351 base pairs encoding 116 amino acids with a theoretical molecular weight of 12.9 kDa, was named as SjMP13 which had 96% of homology with the miracidia associated protein of Schistosoma japonicum. It was confirmed by PCR, double enzyme digestion and DNA sequencing that the recombinant plasmid pET32a （ ＋ ） -SjMP13 was constructed successfully, and expressed in E. coli BL21 （DE3）. Highly purified recombinant protein was prepared by affinity chromatography. The recombinant protein re-SjMP13 could be recognized by the saliva and serum of the schistosomiasis patients.

关 键 词：日本血吸虫 唾液 SjMP13 克隆 表达 

分 类 号：Q78[生物学—分子生物学] S852.735[农业科学—基础兽医学]