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结核分枝杆菌 SWLPK 株的基因组序列和分析。

Genome sequence and analysis of Mycobacterium tuberculosis strain SWLPK.

机构信息

Genomics and Computational Biology Laboratory, Department of Biosciences, COMSATS University Islamabad, Sahiwal Campus, COMSATS Road off GT Road, Sahiwal, Pakistan.

Genomics and Computational Biology Laboratory, Department of Biosciences, COMSATS University Islamabad, Sahiwal Campus, COMSATS Road off GT Road, Sahiwal, Pakistan.

出版信息

J Glob Antimicrob Resist. 2018 Jun;13:211-213. doi: 10.1016/j.jgar.2018.04.014. Epub 2018 May 12.

Abstract

OBJECTIVES

Multidrug-resistant Mycobacterium tuberculosis poses a global threat, particularly in developing countries such as Pakistan. Genome sequencing, comparative genomic analysis and drug resistance gene analysis could be beneficial for understanding and monitoring M. tuberculosis disease severity in Pakistan by elucidating the biology of M. tuberculosis.

METHODS

Here the draft genome of M. tuberculosis strain SWLPK was sequenced, assembled and annotated using an Illumina MiSeq system. De novo genomic assembly was conducted using Geneious Pro™ v.10. The assembled genome of strain SWLPK was annotated using the Rapid Annotation using Subsystem Technology (RAST) server, tRNAscan-SE 1.21 and RNAmmer v.1.2, which provide high-quality functional annotation.

RESULTS

Mycobacterium tuberculosis strain SWLPK yielded an average read depth of 68.5-fold, which covered 97% of the genome of reference strain H37Rv. The genome contains 4305 protein-coding genes, including key drug resistance and virulence-associated genes such as type seven secretion systems. Additionally, it has a 65.6% GC content and contains 48 RNAs and 12 contigs. We determined that all proteins encoded by this strain contain conserved domains, except OxyR, which is associated with first-line antituberculosis drugs such as ethambutol, rifampicin, streptomycin, pyrazinamide and isoniazid.

CONCLUSIONS

This genome sequence provides information regarding the drug resistance genes and virulence propensity of M. tuberculosis strain SWLPK. Strain SWLPK appears to be multidrug-resistant, similar to the Beijing genotype, as it clusters in the same group. These findings will pave the way for genomic characterisation, which will provide further insights into adaptation and evolution in human hosts by transcriptome studies and gene manipulation.

摘要

目的

耐多药结核分枝杆菌对全球构成威胁,尤其是在巴基斯坦等发展中国家。通过阐明结核分枝杆菌的生物学特性,对结核分枝杆菌进行基因组测序、比较基因组分析和耐药基因分析,有助于了解和监测巴基斯坦结核分枝杆菌病的严重程度。

方法

本研究采用 Illumina MiSeq 系统对结核分枝杆菌 SWLPK 株的基因组进行测序、组装和注释。利用 Geneious ProTM v.10 进行从头基因组组装。利用 Rapid Annotation using Subsystem Technology(RAST)服务器、tRNAscan-SE 1.21 和 RNAmmer v.1.2 对 SWLPK 株的组装基因组进行注释,这些工具可提供高质量的功能注释。

结果

结核分枝杆菌 SWLPK 株的平均测序深度为 68.5 倍,覆盖了参考株 H37Rv 基因组的 97%。该基因组包含 4305 个编码蛋白的基因,包括与一线抗结核药物如乙胺丁醇、利福平、链霉素、吡嗪酰胺和异烟肼相关的七型分泌系统等关键耐药和毒力相关基因。此外,其 GC 含量为 65.6%,含有 48 个 RNA 和 12 个连续序列。我们确定该菌株编码的所有蛋白质都含有保守结构域,除了与一线抗结核药物如乙胺丁醇、利福平、链霉素、吡嗪酰胺和异烟肼相关的 OxyR 外。

结论

该基因组序列提供了有关结核分枝杆菌 SWLPK 株耐药基因和毒力倾向的信息。SWLPK 株似乎与北京基因型一样是多药耐药的,因为它与同一组聚类。这些发现将为基因组特征提供信息,通过转录组研究和基因操作,进一步了解在人类宿主中的适应性和进化。

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