A型魏氏梭菌α毒素氨基端PLC结构分析与生物学活性鉴定

发布时间：2018-07-10 17:13

本文选题：A型魏氏梭菌 + α毒素PLC基因　；参考：《浙江农业学报》2017年02期

【摘要】：利用PCR扩增技术克隆A型魏氏梭菌α毒素氨基端的PLC1-250基因,构建含PLC1-250基因表达质粒的BL21(DE3)(p N-PLC1-250)重组菌株,序列分析和酶切鉴定证实构建的p N-PLC1-250重组质粒含有目的基因且基因序列与阅读框架均正确。SDS-PAGE分析表明,PLC1-250蛋白表达量占菌体总蛋白相对含量的18.76%。利用SOPMA法预测PLC1-250蛋白分子的二级结构,且同源模建了其3D结构,结果表明,PLC1-250蛋白分子的二级结构主要为α螺旋和无规则卷曲,三级结构与α毒素相类似。此外,还对其生物学活性进行了鉴定,可为进一步探索α毒素作用的分子机制,以及其分子结构与生物学功能的关系奠定基础。
[Abstract]:PLC1-250 gene was cloned from the amino terminal of 伪 toxin of Clostridium Wei by PCR, and the recombinant strain BL21 (DE3) (p N-PLC1-250) containing PLC1-250 gene expression plasmid was constructed. Sequence analysis and restriction endonuclease analysis confirmed that the recombinant plasmid pN-PLC1-250 contained the target gene and the gene sequence and reading frame were correct. SDS-PAGE analysis showed that the protein expression of pN-PLC1-250 accounted for 18.76% of the relative content of the total cell protein. The secondary structure of PLC1-250 protein molecule was predicted by SOPMA method, and its 3D structure was constructed by homologous model. The results showed that the secondary structure of PLC1-250 protein molecule was mainly 伪 helix and irregular curl, and the tertiary structure was similar to 伪 toxin. In addition, the biological activity of 伪 -toxin was identified, which could lay a foundation for further exploring the molecular mechanism of 伪 -toxin and the relationship between its molecular structure and biological function.
【作者单位】：吉林化工学院生物与食品工程学院;韶关学院英东生命科学院粤北生猪生产及疫病防控协同创新发展中心;吉林农业大学动物科学技术学院;
【基金】：吉林省教育厅“十三五”科学研究规划项目(2016138)
【分类号】：R378

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