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CRISPR-Cas 系统在 中的系统发生分布。

Phylogenetic Distribution of CRISPR-Cas Systems in .

机构信息

Institute of Microbiology and Immunology, School of Life Science, National Yang Ming Chiao Tung University, Taipei, Taiwan.

Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospitalgrid.413801.f, Taoyuan, Taiwan.

出版信息

Microbiol Spectr. 2021 Dec 22;9(3):e0124721. doi: 10.1128/spectrum.01247-21. Epub 2021 Dec 1.

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPRs) and CRISPR-associated (Cas) genes (CRISPR-Cas) are present in many bacterial genomes with functions beyond adaptive immunity. We aimed to characterize the CRISPR-Cas system in the pathogenic Gram-positive bacterium Staphylococcus lugdunensis and determine its association with sequence types (STs) determined by multilocus sequence typing (MLST) and oxacillin susceptibility. Primers were designed to detect and sequence types IIIA and IIC CRISPR-Cas in 199 isolates. MLST and oxacillin susceptibility tests were also performed on the isolates. We found that 84 isolates had type IIIA CRISPR-Cas, while 46 had type IIC. The results showed a strong association between STs and CRISPR-Cas types. The ST1, ST6, ST12, and ST15 isolates had type IIIA CRISPR-Cas systems, and the ST4, ST27, and ST29 isolates had type IIC CRISPR-Cas. Interestingly, of 83 isolates containing type IIIA CRISPR-Cas, 17 (20.5%) were oxacillin-resistant (ORSL), and all of these ORSL isolates belonged to ST6 cluster 1. Moreover, spacers 23 and 21 were found in 16 and 17 ORSL isolates, respectively. In contrast, all 46 isolates with type IIC CRISPR-Cas were susceptible to oxacillin. Our results showed that 41.3% of CRISPR-Cas IIIA spacers were homologous to plasmids and 20.2% were homologous to phages. However, in type IIC CRISPR-Cas, 11.8% and 39.9% of spacers showed sequence homology with plasmids and phages, respectively. In conclusion, we found that the distribution and composition of the CRISPR-Cas system in was associated with STs and oxacillin susceptibility. CRISPR-Cas systems have been characterized as playing several biological roles in many bacterial genomes. Moreover, CRISPR-Cas systems are useful for epidemiological, diagnostic, and evolutionary studies of pathogenic bacteria. However, the characteristics of CRISPR-Cas systems in Staphylococcus lugdunensis have been rarely reported. In this study, we revealed that type IIIA CRISPR-Cas was dominant in isolates, followed by type IIC CRISPR-Cas. Moreover, the composition of CRISPR-Cas spacers was strongly associated with multilocus sequence typing and oxacillin susceptibility of . These results advance our understanding of the evolution of CRISPR-Cas systems; however, the biological functions of CRISPR-Cas systems in remain to be further characterized.

摘要

簇状规律间隔短回文重复序列 (CRISPRs) 和 CRISPR 相关 (Cas) 基因 (CRISPR-Cas) 存在于许多细菌基因组中,具有适应免疫以外的功能。我们旨在描述致病性革兰氏阳性细菌金黄色葡萄球菌 (Staphylococcus lugdunensis) 中的 CRISPR-Cas 系统,并确定其与多位点序列分型 (MLST) 和耐苯唑西林确定的序列类型 (ST) 之间的关联。设计了引物来检测和测序 199 株分离株的 IIIA 和 IIC 型 CRISPR-Cas。还对分离株进行了 MLST 和耐苯唑西林测试。我们发现 84 株分离株具有 IIIA 型 CRISPR-Cas,而 46 株分离株具有 IIC 型 CRISPR-Cas。结果表明 ST 与 CRISPR-Cas 类型之间存在很强的关联。ST1、ST6、ST12 和 ST15 分离株具有 IIIA 型 CRISPR-Cas 系统,而 ST4、ST27 和 ST29 分离株具有 IIC 型 CRISPR-Cas。有趣的是,在含有 IIIA 型 CRISPR-Cas 的 83 株分离株中,有 17 株 (20.5%) 对苯唑西林耐药 (ORSL),所有这些 ORSL 分离株均属于 ST6 簇 1。此外,在 16 株和 17 株 ORSL 分离株中分别发现了间隔物 23 和 21。相比之下,所有 46 株具有 IIC 型 CRISPR-Cas 的分离株均对苯唑西林敏感。我们的结果表明,41.3%的 CRISPR-Cas IIIA 间隔物与质粒同源,20.2%与噬菌体同源。然而,在 IIC 型 CRISPR-Cas 中,分别有 11.8%和 39.9%的间隔物与质粒和噬菌体具有序列同源性。总之,我们发现 中 CRISPR-Cas 系统的分布和组成与 ST 和耐苯唑西林有关。CRISPR-Cas 系统已被描述为在许多细菌基因组中发挥多种生物学作用。此外,CRISPR-Cas 系统可用于致病性细菌的流行病学、诊断和进化研究。然而,金黄色葡萄球菌中 CRISPR-Cas 系统的特征很少被报道。在这项研究中,我们揭示了 IIIA 型 CRISPR-Cas 在 分离株中占主导地位,其次是 IIC 型 CRISPR-Cas。此外,CRISPR-Cas 间隔物的组成与多位点序列分型和 对苯唑西林的敏感性密切相关。这些结果加深了我们对 CRISPR-Cas 系统进化的理解;然而,CRISPR-Cas 系统在 中的生物学功能仍有待进一步表征。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd88/8635126/bbcfdf3a5ace/spectrum.01247-21-f001.jpg

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