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猪链球菌2型核糖核酸酶Ⅲ的克隆表达及催化特性分析

孙文 张永毅 陆畅 朱德文 尹媛媛 徐泉明 陈叶

畜牧与兽医2024,Vol.56Issue(1):46-55,10.
畜牧与兽医2024,Vol.56Issue(1):46-55,10.

猪链球菌2型核糖核酸酶Ⅲ的克隆表达及催化特性分析

Cloning,expression and catalytic properties of ribonuclease Ⅲ from Streptococcus suis serotype 2

孙文 1张永毅 1陆畅 1朱德文 1尹媛媛 1徐泉明 2陈叶1

作者信息

  • 1. 福建农林大学动物科学学院(蜂学学院),闽台动物病原生物学福建省高校重点实验室,福建 福州 350002
  • 2. 福建农林大学动物科学学院(蜂学学院),闽台动物病原生物学福建省高校重点实验室,福建 福州 350002||福建警察学院,福建 福州 350007
  • 折叠

摘要

Abstract

In order to identify the catalytic properties of RNase Ⅲ Streptococcus suis serotype 2 RNase Ⅲ(SS-RNase Ⅲ),we used Esche-richia coli BL21(DE3)to heterologously express RNase Ⅲ of Streptococcus suis serotype 2,and the bacterial RNase Ⅲ was then isolated and purified by nickel column affinity chromatography.Next,various catalytic properties of SS-RNase Ⅲ were analyzed.The results were that heterologous expression showed most of the recombinant protein RNase Ⅲ to be soluble,that the catalytic properties showed RNase Ⅲ mainly relying on Mg2+or Mn2+to perform catalytic functions with different catalytic activities under different ion concentrations and pH environments,that substrate specific analysis showed RNase Ⅲ to specifically cleave double stranded RNA with a length of 30bp,with good specificity for endogenous rnc mRNA and 5S rRNA.The above results indicated that SS-RNase Ⅲ had unique catalytic properties,which provided theoreti-cal basis for a deeper understanding of how SS-RNase Ⅲ mediates the pathogenesis of Streptococcus suis serotype 2.

关键词

猪链球菌2型/核糖核酸酶Ⅲ/原核表达/催化特性

Key words

Streptococcus suis serotype 2/ribonuclease Ⅲ/prokaryotic expression/catalytic properties

分类

农业科技

引用本文复制引用

孙文,张永毅,陆畅,朱德文,尹媛媛,徐泉明,陈叶..猪链球菌2型核糖核酸酶Ⅲ的克隆表达及催化特性分析[J].畜牧与兽医,2024,56(1):46-55,10.

基金项目

福建省自然科学基金项目(2023J01272) (2023J01272)

福建农林大学科技创新专项(CXZX2020061A) (CXZX2020061A)

畜牧与兽医

OA北大核心CSTPCD

0529-5130

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