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金黄色葡萄球菌RNAⅢ敲除株对γ-溶血素表达调控的影响
Effect of deletion of
Staphylococcus aureus RNAⅢ on γ-hemolysin expression- 第三军医大学学报 2020年42卷第1期 页码：45-49
- 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：
[胡晓梅，E-mail:hxmay2008@163.com HU Xiaomei, E-mail:hxmay2008@163.com
- 基金信息：国家自然科学基金(31570127);国家自然科学基金(31470241)
DOI：10.16016/j.1000-5404.201908007
中图分类号： R378.11;R394-33;R394.2收稿：2019-08-01，
修回：2019-08-23，
纸质出版：2020-01-15
- 稿件说明：
移动端阅览
谭利, 王轶男, 陈江, 等. 金黄色葡萄球菌RNAⅢ敲除株对γ-溶血素表达调控的影响[J]. 第三军医大学学报, 2020,42(1):45-49.
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.谭利, 王轶男, 陈江, 等. 金黄色葡萄球菌RNAⅢ敲除株对γ-溶血素表达调控的影响[J]. 第三军医大学学报, 2020,42(1):45-49. DOI： 10.16016/j.1000-5404.201908007.
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
2
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
2
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
2
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
2
RNAⅢ敲除可显著影响金葡菌γ-溶血素的表达。
Abstract
Objective
2
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
2
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
2
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
2
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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