猪囊尾蚴抗原基因GP50的表达及鉴定  

Expression and identification on the antigen GP50 gene of Cysticercus cellulosae

作　　者：周天祥[1] 汪学龙[2] 张衍兴[1] 郑胜生[2] 王旭旭[1] 丁言荣[1] 

机构地区：[1]安徽省淮北职业技术学院,淮北235000 [2]安徽医科大学病原生物学教研室

出　　处：《热带病与寄生虫学》2007年第2期85-87,共3页Journal of Tropical Diseases and Parasitology

基　　金：安徽省淮北职业技术学院自然科学基金资助项目

摘　　要：目的为了寻找猪囊尾蚴病新的免疫学候选诊断分子,克隆猪囊尾蚴抗原GP50编码基因,并进行表达、鉴定。方法设计合成引物,用PCR法从猪囊尾蚴cDNA文库中扩增出猪囊尾蚴抗原GP50基因编码序列,将其克隆入pGEM-T载体,然后在真核表达载体pBKCMV中亚克隆,用IPTG诱导表达,SDS-PAGE和Westernblot观察表达结果。结果RT-PCR法扩增出一条大小约897bp的特异性片段,克隆质粒pGEM-GP50和真核表达质粒pBKCMV作BamHⅠ和XhoⅠ双酶切和以重组质粒为模板进行PCR扩增,均可获得一条与PCR产物一致的DNA片段。诱导表达后,经SDS-PAGE可见一条约36kDa大小的融合蛋白条带,Western blot结果显示其可与猪囊尾蚴病人血清起反应。结论本实验成功地克隆了猪囊尾蚴抗原GP50编码基因,并在原核细胞中进行了表达及鉴定,为进一步的免疫诊断研究奠定了基础。Objective In order to probe a new candidate molecular for the immunodiagnosis of cysticercosis ceUulosae,the antigen GP50 gene of Cysticercus cellulosae was cloned, expressed and identified in the prokaryotic cell. Methods The specifc primers were designed and synthesized. The DNA fragment encoding GP50 was amplified by PCR from the cDNA library of Cysticercus cellulosae. The PCR products were cloned into pGEM-T vector, then the antigen GPS0 gene was subcloned into pBK-CMV expression vector. IPTG was added to induce fusion expression of fused GPS0. The expression was identified by SDS- PAGE and Western blot. Results The RT-PCR amplified product was 897bp in size. The size of pGEM- T-GP50 and pBK-CMV-GP50 digested with BamH I and Xho I was identical to the length of the PCR generated products. After inducible expression, one fusion protein band with 36kDa in size was identified by SDS-PAGE. The fusion protein could be recognized by the sera of patients infected with cysticercosis, Condusion The cDNA encoding antigen GP50 gene of Cysticercus cellulosae is cloned successfully, and it can be expressed and identificated in the prokaryotic cell, which provides the basis for further study on immunodiagnosis of cysticercosis,

关 键 词：猪囊尾蚴 抗原 基因克隆 表达 

分 类 号：R532.33[医药卫生—内科学]