本文選題：A型魏氏梭菌 + α毒素PLC基因��； 參考：《浙江農(nóng)業(yè)學(xué)報》2017年02期

【摘要】：利用PCR擴(kuò)增技術(shù)克隆A型魏氏梭菌α毒素氨基端的PLC1-250基因,構(gòu)建含PLC1-250基因表達(dá)質(zhì)粒的BL21(DE3)(p N-PLC1-250)重組菌株,序列分析和酶切鑒定證實構(gòu)建的p N-PLC1-250重組質(zhì)粒含有目的基因且基因序列與閱讀框架均正確。SDS-PAGE分析表明,PLC1-250蛋白表達(dá)量占菌體總蛋白相對含量的18.76%。利用SOPMA法預(yù)測PLC1-250蛋白分子的二級結(jié)構(gòu),且同源模建了其3D結(jié)構(gòu),結(jié)果表明,PLC1-250蛋白分子的二級結(jié)構(gòu)主要為α螺旋和無規(guī)則卷曲,三級結(jié)構(gòu)與α毒素相類似。此外,還對其生物學(xué)活性進(jìn)行了鑒定,可為進(jìn)一步探索α毒素作用的分子機(jī)制,以及其分子結(jié)構(gòu)與生物學(xué)功能的關(guān)系奠定基礎(chǔ)。
[Abstract]:PLC1-250 gene was cloned from the amino terminal of 偽 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 偽 helix and irregular curl, and the tertiary structure was similar to 偽 toxin. In addition, the biological activity of 偽 -toxin was identified, which could lay a foundation for further exploring the molecular mechanism of 偽 -toxin and the relationship between its molecular structure and biological function.
【作者單位】： 吉林化工學(xué)院生物與食品工程學(xué)院;韶關(guān)學(xué)院英東生命科學(xué)院粵北生豬生產(chǎn)及疫病防控協(xié)同創(chuàng)新發(fā)展中心;吉林農(nóng)業(yè)大學(xué)動物科學(xué)技術(shù)學(xué)院;
【基金】：吉林省教育廳“十三五”科學(xué)研究規(guī)劃項目(2016138)
【分類號】：R378

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本文編號：2114059