魏氏梭菌α毒素基因的克隆表达及其间接ELISA方法的建立

Cloning and expression of α-toxin gene of Clostridium welchii and establishment of indirect ELISA

对GenBank上发表的魏氏梭菌α毒素基因的氨基酸序列进行了二级结构、疏水性、抗原决定簇、跨膜区以及抗原性分析,在此基础上,对α毒素基因130～966nt位的837bp长的适合原核表达的区域进行PCR扩增、克隆、核酸序列测定和生物信息学分析。同时,建立了检测魏氏梭菌α毒素抗体的间接ELISA方法。

According to the published genome sequences of α-toxin gene of Clostridium welchii on GenBank, the study analysed the secondary structure of amino acid sequence, hydrophobicity, antigenic determinant, transmembrane region and antigenic region of α-toxin gene of Clostridiurn welchii. Therefore, the study choosed the region of α-toxin gene which was 837 bp in length between 130- 966 nt and expressed compatibly in prokaryotic ceils for PCR amplification,cloning, sequencing and bioinformaties analysis. Meanwhile, an indirect ELISA method detecting antibody against α-toxin of Clostridium welchii was estabished.