大肠杆菌不耐热肠毒素B亚单位与产气荚膜梭菌β_1、β_2毒素融合基因的构建及表达

Construction and expression of the fusion gene of beta1-toxin gene and beta2-toxin gene from Clostridium perfringens with Escherichia coli heat-labile enterotoxin B subunit

为了构建含大肠杆菌不耐热肠毒素B亚单位(LTB)与产气荚膜梭菌β1、β2毒素融合基因的表达菌株,试验将亚克隆LTB基因融合到β2-β1毒素基因的上游,构建了pET30a-LTB-β2-β1原核表达载体,经IPTG诱导表达,对其表达产物进行SDS-PAGE检测和Western-blot分析。结果表明:重组菌株可以表达LTB-β2-β1融合蛋白,且该融合蛋白可以被相应的抗体识别。

To research the induced expression of the fusion protein by the β1 and β2 - toxins of Clostridium perfringens and E. coli heat - labile enterotoxin B subunit. A 306 bp LTB DNA fragment was amplified with PCR, and inserted the upstream of Beta2 and Betal fusion gene, then transformed into the receptor strain Escherichia coli BL21 （ DE3 ）. Preliminary expression of the LTB - β2 - β1 fusion gene was optimized by inducing IPTG, and its expression products were detected by SDS - PAGE and Western - blot analysis. The results showed that the fusion gene could be expressed and specifically recognized by the corresponding antibodies.