[Role of CRISPR/Cas systems in drugresistance and virulence and the effect of IS600 on the expression of cse2 in Shigella].

微生物学报 Pub Date : 2016-12-04
Lijuan Hong, Bing Zhang, Guangcai Duan, Wenjuan Liang, Yingfang Wang, Shuaiyin Chen, Haiyan Yang, Yuanlin Xi
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Abstract

Objective: To analyze the relationship between CRISPR/Cas system and drug-resistance, virulence. To investigate the effect of IS600 on the expression of CRISPR associated gene cse2 in Shigella.

Methods: CRISPR loci, CRISPR associated gene cse2, drug-resistant genes and virulent genes were detected by PCR in 33 Shigella strains; Trypan Blue counting test was used to detect bacterial virulence; Real-time PCR was used to detect relative mRNA expression of cse2; susceptibilities of Shigella strains were tested by agar diffusion method. Furthermore, we analyzed the relationship between CRISPR loci and drug-resistant genes, virulent genes. The effect of the IS600 on the expression of CRISPR associated gene cse2 was investigated.

Results: The mortality of Hela cells infected by Shigella with CRISPR1 loci was significantly lower (P<0.05) than those infected by Shigella without CRISPR1. The mRNA expression level of cse2 in group with IS600 was significantly (P<0.05) lower than that in group without IS600.

Conclusions: CRISPR loci were widely present in Shigella. Shigella without CRISPR1 has a higher pathogenicity. Due to the insertion of IS600, the mRNA expression level of cse2 was decreased in Shigella.

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[CRISPR/Cas系统在志贺氏菌耐药和毒力中的作用以及IS600对cse2表达的影响]。
目的:分析CRISPR/Cas系统与耐药、毒力的关系。目的探讨IS600对志贺菌中CRISPR相关基因cse2表达的影响。方法:采用PCR检测33株志贺氏菌的CRISPR基因座、CRISPR相关基因cse2、耐药基因和毒力基因;台盼蓝计数法检测细菌毒力;Real-time PCR检测cse2 mRNA相对表达量;用琼脂扩散法检测志贺氏菌的药敏。进一步分析了CRISPR位点与耐药基因、毒力基因的关系。研究了IS600对CRISPR相关基因cse2表达的影响。结果:携带CRISPR1基因座的Hela细胞感染志贺氏菌后死亡率明显降低(p结论:CRISPR基因座在志贺氏菌中广泛存在。没有CRISPR1的志贺氏菌具有更高的致病性。由于IS600的插入,cse2 mRNA在志贺氏菌中的表达水平降低。
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期刊介绍: Acta Microbiologica Sinica(AMS) is a peer-reviewed monthly (one volume per year)international journal,founded in 1953.It covers a wide range of topics in the areas of general and applied microbiology.The journal publishes original papers,reviews in microbiological science,and short communications describing unusual observations. Acta Microbiologica Sinica has been indexed in Index Copernicus (IC),Chemical Abstract (CA),Excerpt Medica Database (EMBASE),AJ of Viniti (Russia),Biological Abstracts (BA),Chinese Science Citation Database (CSCD),China National Knowledge Infrastructure(CNKI),Institute of Scientific and Technical Information of China(ISTIC),Chinese Journal Citation Report(CJCR),Chinese Biological Abstracts,Chinese Pharmaceutical Abstracts,Chinese Medical Abstracts and Chinese Science Abstracts.
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