生物技术进展 ›› 2022, Vol. 12 ›› Issue (2): 313-317.DOI: 10.19586/j.2095-2341.2021.0136

• 研究论文 • 上一篇    

金黄色葡萄球菌肠毒素A、B、C、D、E重组载体的构建及表达

陈威风1(), 崔丽伟1, 常惟丹1, 朱龙佼2, 许文涛2()   

  1. 1.河南牧业经济学院食品与生物工程学院,郑州 450046
    2.中国农业大学营养与健康系,北京 100083
  • 收稿日期:2021-07-15 接受日期:2021-08-24 出版日期:2022-03-25 发布日期:2022-03-25
  • 通讯作者: 许文涛
  • 作者简介:陈威风 E-mail: cwf86@webmail.hzau.edu.cn
  • 基金资助:
    河南牧业经济学院博士启动经费项目(51000961);河南省科技攻关项目(182102110303)

Construction and Expression of Recombinant Vectors of Staphylococcal Enterotoxins A, B, C, D and E

Weifeng CHEN1(), Liwei CUI1, Weidan CHANG1, Longjiao ZHU2, Wentao XU2()   

  1. 1.School of Food and Biological Engineering,Henan University of Animal Husbandry and Economy,Zhengzhou 450046,China
    2.Department of Nutrition and Health,China Agricultural University,Beijing 100083,China
  • Received:2021-07-15 Accepted:2021-08-24 Online:2022-03-25 Published:2022-03-25
  • Contact: Wentao XU

摘要:

金黄色葡萄球菌肠毒素(Staphylococcal enterotoxins,SEs)是一组结构毒力相似、血清型不同的可溶性小分子蛋白质,平均分子质量为26~30 kD,是引起细菌性食物中毒及肠胃炎的主要因素之一。为了制备金黄色葡萄球菌肠毒素的纯品,首先合成了SEA、SEB、SEC、SED和SEE的基因序列,然后构建了5种肠毒素的原核表达载体,分别转入BL21(DE3)细胞中进行诱导表达。通过SDS-PAGE和Western blot验证,5种肠毒素蛋白均被成功表达,并且在较低的诱导温度(16 ℃)获得一定量的可溶性蛋白。成功制备了5种金黄色葡萄球菌肠毒素的可溶性蛋白,为今后更好地解决因SEs引起的食品安全问题奠定了基础。

关键词: 金黄色葡萄球菌肠毒素, 食物中毒, 原核表达载体, 可溶性蛋白

Abstract:

As one of the main factors causing bacterial food poisoning and gastroenteritis, Staphylococcal enterotoxins (SEs) is a group of soluble small-molecule proteins with similar structural toxicity and different serotypes with the average molecular mass of 26~30 kD. To prepare pure products of SEs, the gene sequence of SEA, SEB, SEC, SED and SEE were synthesized, then the proraryotic expression vectors of five enterotoxins were constructed and transferred into BL21(DE3) cells for expression. SDS-PAGE and Western blot showed that the five enterotoxin proteins were successfully expressed, and a amount of soluble proteins could be obtained at a lower induction temperature (16 ℃). In this study, five soluble proteins of Staphylococcal enterotoxin were successfully prepared, which laid a foundation for better solving the food safety problems caused by SEs in the future.

Key words: Staphylococcal enterotoxins, food poisoning, prokaryotic expression vectors, soluble proteins

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