鉴定和分析肺炎克雷伯氏菌耐抗生素菌株基因组中的 CRISPR/Cas 系统结构

L. A. Stepanenko, B. G. Sukhov, T. V. Kon’kova, V. V. Bedinskaya, N. V. Klushina, V. I. Zlobin
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引用次数: 0

摘要

背景。肺炎克雷伯氏菌属于机会性细菌,可对抗生素形成多重耐药性,并通过水平基因转移将其传播给各类细菌。这些研究考察了保护细菌免受外来 DNA 侵害的 CRISPR/Cas 系统的结构和功能多样性。他们以肺炎克雷伯氏菌的抗生素耐药菌株为例进行分析,将证明这些菌株对某些噬菌体的耐药性,从而有可能开发出通过创建靶向噬菌体疗法来治疗由这些微生物引起的复杂传染性疾病的方法。以肺炎克雷伯氏菌的抗生素耐药菌株为例,对已确定的 CRISPR/Cas 系统的结构成分进行生物信息学分析,通过 CRISPR 盒式间隔筛选噬菌体。文章分析了 29 个肺炎克雷伯菌的全基因组序列,确定了其中 CRISPR/Cas 系统和抗生素耐药基因的结构(根据 NCBI)。为实现这一目标,利用软件建模方法对 Cas 基因和 CRISPR 盒进行了搜索,并给出了它们的结构和功能特征。利用生物信息学搜索算法,在抗生素耐药菌株的基因组中发现了功能活跃的 CRISPR/Cas 系统,这些系统含有一个或两个 CRISPR 盒,属于 I 型亚型 IE。鉴定出的耐药菌株群具有相同的 CRISPR 盒的间隔成分。系统进化分析证实了它们的共同起源。通过分析 CRISPR 盒的间隔序列,确定了同属肠杆菌科的克雷伯氏菌属、沙门氏菌属细菌的噬菌体多样性谱。因此,获得了肺炎克雷伯菌株具有抗生素耐药性的 CRISPR 系统作用所针对的噬菌体的信息。对肺炎克雷伯氏菌耐抗生素菌株的 CRISPR/Cas 系统的功能和结构特征进行分析,可以获得有关其进化史以及其作用所针对的噬菌体的信息,即有关其噬菌体耐药性的信息。本研究采用的方法可进一步作为创建个性化噬菌体疗法的基础
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Identification and analysis of CRISPR/Cas systems structures in the genomes of antibiotic-resistant strains of Klebsiella pneumoniae
Background. Klebsiella pneumoniae belongs to a group of opportunistic bacteria that can form multiple resistance to antibiotics and transmit it to various types of bacteria through horizontal gene transfer. These studies examine the structural and functional diversity of CRISPR/Cas systems that protect bacteria from foreign DNA. Their analysis using the example of antibiotic-resistant strains of Klebsiella pneumoniae will demonstrate their resistance to certain bacteriophages, which will make it possible to develop approaches to the treatment of complex infectious diseases caused by these microorganisms by creating targeted phage therapy.The aim. To perform a bioinformatics analysis of the identified structural components of CRISPR/Cas systems for screening bacteriophages through CRISPR cassette spacers using the example of antibiotic-resistant strains of Klebsiella pneumoniae.Materials and methods. The article analyzed 29 full-genome sequences of Klebsiella pneumoniae, in the genome of which the structures of CRISPR/Cas systems and  antibiotic resistance genes were determined (according to NCBI). To achieve this  goal, using software modeling methods, a search was made for Cas genes and CRISPR cassettes, and their structural and functional characteristics were given.Results. Using bioinformatic search algorithms in the genome of antibiotic-resistant strains, functionally active CRISPR/Cas systems with the presence of one or two CRISPR cassettes and belonging to Type  I Subtype  IE were identified. Groups of resistant strains with identical spacer composition of CRISPR cassettes have been identified. A phylogenetic analysis was carried out confirming their common origin. By analyzing the spacer sequences of CRISPR cassettes, the spectrum of diversity of phages of bacteria of the genus Klebsiella, Salmonella, belonging to the same family Enterobacteriaceae, was determined. Thus, information was obtained about the bacteriophages that are targeted by the action of CRISPR systems of Klebsiella pneumoniae strains that have antibiotic resistance.Conclusions. Analysis of the functional and structural features of the CRISPR/Cas systems of antibiotic resistant Klebsiella pneumoniae strains made it possible to obtain information about their evolutionary history and about the bacteriophages against which their action is directed, that is, about their phage resistance. The approach used in this study may further serve as the basis for the creation of personalized phage therapy
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