三种方法对日本血吸虫抗原cDNA克隆的筛选和鉴定  

Screening and identification of antigen cDNA clones of Schistosoma japonica by three methods

作　　者：沈定文[1] 罗金萍[1] 詹希美[2] 刘启文[2] 王燕妮[2] 

机构地区：[1]咸宁医学院寄生虫学教研室,咸宁437100 [2]中山医科大学寄生虫学教研室,广州510089

出　　处：《华中医学杂志》1997年第4期172-173,共2页Central China Medical Journal

基　　金：美国中华医学基金会(CMB)资助

摘　　要：利用融源表达、平板裂解法和PCR扩增三种不同方法分别对日本血吸虫抗原cDNA基因进行鉴定和分析,8个免疫筛选阳性克隆均能在大肠杆菌中以融合蛋白的形式表达,表达蛋白分子量为140～150kDa,抗原cDNA基因经限制性内切酶EcoRI酶解后,琼脂糖凝胶电泳显示其大小为700～900bp,PCR能扩增出特异性条带。结果表明,上述三种方法能从蛋白质和核酸水平有效地鉴定血吸虫抗原cDNA基因。Schistosoma japonica antigen cDNA clones were identified by lysogenic expression,flat lyiic method and PCR amplification. All 8 positive clones immunologically screened could be expressed in E- coli in the form of fusion proteins with the molecular weight being about 140 to 150 kDa. The positive cDNA genes were digested by restriction endonuclease EcoRI,then the agarose gel electrophoresis revealed the size of them being 700 to 900 bp. The antigen cDNA genes could be specifically amplified by PCR. The results indicated that schistosoma japonica antigen cDNA genes could be identified effectively by means of these three methods.

关 键 词：日本血吸虫 抗原 CDNA 筛选 鉴定 

分 类 号：R383.24[医药卫生—医学寄生虫学]