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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。
Three different methods were used to identify and analyze the cDNA of Schistosoma japonicum antigen, respectively. The eight immunoscreening positive clones were all expressed as fusion protein in E. coli, and the protein molecular weight Is 140-150 kDa. After the cDNA gene of the antigen was digested with restriction enzyme EcoRI, the size of the antigen cDNA was 700-900 bp by agarose gel electrophoresis, and the specific band was amplified by PCR. The results show that these three methods can effectively identify the schistosome antigen cDNA gene from the protein and nucleic acid levels.