A型魏氏梭菌α毒素氨基端plc基因的生物学活性探究

Biological Activity Exploration of plc Gene at α-toxin Amino-Terminal from Clostridium welchii Type A

为了探究α毒素氨基端及其生物学功能的关系,本研究利用PCR技术克隆了魏氏梭菌α毒素氨基端基因PLC1-250,构建了含PLC1-250基因重组质粒的BL21(DE3)(p N-PLC1-250)重组表达菌株。根据序列分析和酶切鉴定的结果,证实了重组质粒p N-PLC1-250含有PLC1-250基因,且其开放阅读框架和基因序列均正确。经过SDS-PAGE分析,结果表明PLC1-250蛋白表达量约占菌体总蛋白相对含量的18.76%。对其生物学活性的研究表明,PLC1-250蛋白与α毒素一样具有磷脂酶C活性。研究为进一步揭示α毒素的分子作用机制提供了有力的支持。

In order to explore the relationship between the amino-terminal of α-toxin and its biological function,this research used PCR technology to clone PLC1-250, the amino-terminal gene of α-toxin from Clostridium welchii, constructing the recombinant expression strains BL21（DE3）（p N-PLC1-250） which contained gene recombinant plasmid PLC1-250. According to the results of sequence analysis and enzyme identification, the recombinant plasmid p N-PLC1-250 was proved to contain PLC1-250 gene, of which the ORF（open reading frame）and sequence were both correct. The result of SDS-PAGE analysis showed that the expression level of PLC1-250 protein in the relative amount of total bacterial protein was about 18.76%. According to biological activity research,PLC1-250 protein was proved to have the activity of phospholipase C, which was the same as α-toxin. This research provided powerful support for the further study in revealing the molecular mechanism of α-toxin.