A型产气荚膜梭菌噬菌体裂解酶Cp51的原核表达及活性检测

发布时间：2018-01-19 15:28

本文关键词： 产气荚膜梭菌噬菌体裂解酶蛋白表达杀菌活性　出处：《华南农业大学学报》2017年05期 　论文类型：期刊论文

【摘要】：【目的】原核表达A型产气荚膜梭菌噬菌体裂解酶Cp51并研究其对A型产气荚膜梭菌Clostridium perfringens的体外抗菌活性。【方法】合成噬菌体裂解酶Cp51基因,并构建了pET-32a-Cp51原核表达载体,转化到大肠埃希菌Escherichia coli BL21(DE3),经终浓度为0.5 mmol·L~(-1)的IPTG诱导以及镍柱纯化后,获得了可溶性的Cp51重组蛋白;用比浊法检测Cp51重组蛋白的杀菌活性。【结果】噬菌体裂解酶Cp51重组蛋白能够有效降低7株A型产气荚膜梭菌的浊度,裂解酶质量浓度在5μg·m L~(-1)以上、作用30 min对A型产气荚膜梭菌的杀菌率可达到99.99%以上,而对其他种类细菌无杀菌效果。【结论】A型产气荚膜梭菌噬菌体裂解酶Cp51重组蛋白对A型产气荚膜梭菌有较强的体外杀菌活性和特异性,研究结果为后续裂解酶Cp51的临床应用奠定基础。
[Abstract]:[objective] to express the phage lyase Cp51 of Clostridium perfringens type A in prokaryotic expression and to study its effect on Clostridium of Clostridium perfringens type A. In vitro antibacterial activity of perfringens. [methods] Phage lyase Cp51 gene was synthesized. The prokaryotic expression vector of pET-32a-Cp51 was constructed and transformed into Escherichia coli Escherichia coli BL21 (DE3). The soluble recombinant Cp51 protein was obtained by IPTG induction and nickel column purification at the final concentration of 0.5 mmol 路L ~ (-1). The bactericidal activity of Cp51 recombinant protein was detected by turbidimetry. [results] the bacteriophage lyase Cp51 recombinant protein could effectively reduce the turbidity of 7 strains of Clostridium perfringens A. The bactericidal rate of Clostridium perfringens type A was over 99.99% when the lyase concentration was above 5 渭 g 路mL ~ (-1) and exposed for 30 min. Conclusion the recombinant protein of bacteriophage lyase Cp51 of Clostridium perfringens type A has strong bactericidal activity and specificity against Clostridium perfringens type A. The results laid a foundation for clinical application of Cp51.
【作者单位】：华南农业大学兽医学院/广东省动物源性人兽共患病预防与控制重点实验室;
【基金】：广东省科技计划项目(2014A020208099)
【分类号】：S852.61
【正文快照】： 优先出版网址:http://kns.cnki.net/kcms/detail/44.1110.s.20170714.0855.008.html产气荚膜梭菌Clostridium perfringens又名魏氏梭菌C.welchii,是一种重要的人畜共患病原菌[1]。产气荚膜梭菌为两端钝圆厌氧性菌,呈粗杆状,单个或成对存在,是一种能产芽孢的革兰阳性菌,在人和

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