[1]谭利,李思瑞,姜北,等.金黄色葡萄球菌酚溶调控蛋白α小肽敲除对自溶素AtlA蛋白表达的影响[J].第三军医大学学报,2017,39(21):2066-2071.
 TAN Li,LI Sirui,JIANG Bei,et al.Effect of PSMα knockout on autolysin AtlA expression in Staphylococcus aureus[J].J Third Mil Med Univ,2017,39(21):2066-2071.
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金黄色葡萄球菌酚溶调控蛋白α小肽敲除对自溶素AtlA蛋白表达的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
39卷
期数:
2017年第21期
页码:
2066-2071
栏目:
基础医学
出版日期:
2017-11-15

文章信息/Info

Title:
Effect of PSMα knockout on autolysin AtlA expression in Staphylococcus aureus
作者:
谭利李思瑞姜北张骁鹏陈江胡晓梅黎庶
第三军医大学基础医学部微生物学教研室
Author(s):
TAN Li LI Sirui JIANG Bei ZHANG Xiaopeng CHEN Jiang HU Xiaomei LI Shu

Department of Microbiology, College of Basic Medical Sciences, Third Military Medical University, Chongqing, 400038, China

关键词:
金黄色葡萄球菌PSM&alpha小肽自溶素AtlA蛋白
Keywords:
Staphylococcus aureus PSM&alpha peptides autolysin AtlA protein  
分类号:
R378.11; R39433; R394.3
文献标志码:
A
摘要:

目的    利用同源重组技术构建金黄色葡萄球菌Newman psmα基因缺失突变株,探究psmα缺失对金黄色葡萄球菌蛋白表达的影响。方法  分别扩增psmα基因的上下游同源臂,根据同源重组原理,构建pBT2基因打靶载体,利用同源重组技术,构建psmα缺失的Newman突变株。并对野生株和突变株培养上清蛋白进行研究。结果    突变株培养上清中一个相对分子质量约为135×103的蛋白表达水平较野生株明显减少甚至缺失;质谱分析提示该蛋白为金黄色葡萄球菌atlA基因编码的双功能自溶素前体蛋白(AtlA蛋白);实时荧光定量PCR结果显示psmα基因敲除株atlA基因的转录水平较野生株显著降低。结论    psmα缺失后能够降低atlA基因的转录表达水平。

Abstract:

Objective     To construct the phenol-soluble modulin (psmα) locus gene knockout mutant of Staphylococcus aureus (S. aureus) and evaluate the effect of PSMα peptides on the expression of secretory proteins. Methods     The temperature-sensitive S. aureus knock-out vectors were constructed after 2 homologous sequences around the psmα locus were cloned and inserted into pBT2 vector to construct a knockout plasmid pBT2∷psmα. Then the obtained plasmids pBT2∷psmα were transferred into S. aureus strain Newman, and the Newman∷Δpsmα mutant were prepared with homologous recombination. The proteins in the culture supernatants of Newman wild type strain and Newman∷Δpsmα mutant were analyzed with SDS-polyacrylamide gel electrophoresis (SDSPAGE) and mass spectrometry. The transcriptional level of altA gene was detected using quantitative real-time PCR. Results     SDS-PAGE confirmed that there was a band missing at the relative mass weight of 135×103 when compared with the results of the wildtype strain. Mass spectrometry indicated that the protein was dual functionality autolyzed precursor protein (AtlA) in Newman∷Δpsmα mutant. RT-PCR also showed the atlA gene transcription was significantly reduced in the psmα mutant strain. Conclusion     The PSMα knockout in S. aureus reduces the expression of atlA at transcriptional level.
 

参考文献/References:

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更新日期/Last Update: 2017-11-09