Effect of deletion of <italic style="font-style: italic">Staphylococcus aureus</italic> RNAⅢ on γ-hemolysin expression

Li TAN; Yinan WANG; Jiang CHEN; Bei JIANG; Shengpeng YU; Shu LI; Xiaomei HU

doi:10.16016/j.1000-5404.201908007

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Effect of deletion of Staphylococcus aureus RNAⅢ on γ-hemolysin expression

Basic Medicine | 更新时间：2026-01-09

- Effect of deletion of Staphylococcus aureus RNAⅢ on γ-hemolysin expression
- Journal of Third Military Medical University Vol. 42, Issue 1, Pages: 45-49(2020)
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学):基础医学院微生物学教研室
  2.400038 重庆，陆军军医大学(第三军医大学):第一附属医院全军烧伤研究所，创伤、烧伤与复合伤国家重点实验室
  3.400038 重庆，陆军军医大学(第三军医大学):第一附属医院骨科，全军矫形外科中心
- 作者简介：
- 基金信息：
  
  the National Natural Science Foundation of China(31570127);the National Natural Science Foundation of China(31470241)
- DOI：10.16016/j.1000-5404.201908007
  CLC： R378.11;R394-33;R394.2
- Received：01 August 2019，
  
  Revised：2019-08-23，
  
  Published：15 January 2020
- 稿件说明：
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Li TAN, Yinan WANG, Jiang CHEN, et al. Effect of deletion of Staphylococcus aureus RNAⅢ on γ-hemolysin expression[J]. Journal of Third Military Medical University, 2020, 42(1): 45-49.
DOI：

Li TAN, Yinan WANG, Jiang CHEN, et al. Effect of deletion of Staphylococcus aureus RNAⅢ on γ-hemolysin expression[J]. Journal of Third Military Medical University, 2020, 42(1): 45-49. DOI： 10.16016/j.1000-5404.201908007.

摘要

目的

利用同源重组技术构建金黄色葡萄球菌Newman RNAⅢ基因敲除株，探究RNAⅢ敲除对金葡菌γ-溶血素表达调控的影响。

方法

分别扩增RNAⅢ基因的上下游同源臂，根据同源重组原理，构建pBT2基因敲除载体，利用同源重组技术，构建RNAⅢ敲除株。并对野生株和敲除株γ-溶血素的转录水平和蛋白水平进行研究。

结果

实时荧光定量PCR结果显示RNAⅢ敲除株γ-溶血素A/B/C亚基的转录水平较野生株显著降低

Western blot检测结果也显示敲除株γ-溶血素蛋白表达水平较野生株明显减少。

结论

RNAⅢ敲除可显著影响金葡菌γ-溶血素的表达。

Abstract

Objective

To construct a RNAⅢ gene knockout mutant of

Staphylococcus aureus

(

S. aureus

) and evaluate the effect of RNAⅢ on the expression of γ-hemolysin.

Methods

In order to construct temperature-sensitive

S. aureus

knock-out vectors

2 homologous sequences around RNAⅢ were cloned and inserted into pBT2 vector to construct knock-out plasmid pBT2∷RNAⅢ

and then the obtained plasmids RNAⅢ were transferred into

S. aureus

strain Newman

finally Newman∷ΔRNAⅢ mutant were prepared with homologous recombination. The transcriptional level of γ-hemolysin hlgA/B/C was detected by quantitative real-time PCR (RT-PCR) and the protein levels in the culture supernatants of Newman wild type strain and Newman∷ΔRNAⅢ were analyzed with SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis.

Results

Newman∷ΔRNAⅢ mutant was constructed successfully. RT-PCR analysis revealed that the expression of γ-hemolysin in the Newman∷ΔRNAⅢ mutant was significantly decreased as compared to that of Newman wild type strain. Western blotting also showed the transcription level of γ-hemolysin was significantly reduced in the mutant strain.

Conclusion

The knockout of RNAⅢ has a great effect on the expression of γ-hemolysin in

S. aureus

关键词

Keywords

references

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Effect of deletion of Staphylococcus aureus RNAⅢ on γ-hemolysin expression

DOI：10.16016/j.1000-5404.201908007

摘要

Abstract

关键词

Keywords

references