An efficient method for knocking out genes on the virulence plasmid of hypervirulent Klebsiella pneumoniae.

IF 1.5 4区医学 Q4 MICROBIOLOGY

New Microbiologica Pub Date : 2023-05-01

Caiqin Zi, Si Yang, Xiaofang Fu, Weiqi Wang, Yi Luo, Jing Zhang, Wentao Guo, Heping Wang, Li Li, Xinyun Liang, Na Mi, Tingting Zhi, Zuguo Zhao

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引用次数: 0

Abstract

Currently, the infection of hypervirulent Klebsiella pneumoniae (hvKp) is becoming increasingly serious and the virulent mechanisms of hvKp are still not very clear. An effective gene-editing method for genes on hvKp virulence plasmid can help us reveal related virulent mechanisms. There are a few reports focusing on the methods mentioned above, however with certain limitations. In this work, we first constructed the pRE112-basing recombinant suicide plasmid to knock out or replace the genes in the hvKp virulence plasmid based on the principle of homology recombination. Results showed that the target virulent genes iucA, iucB, iroB, and rmpA2 on the hvKp virulence plasmid were scarlessly knocked out or replaced by marker genes, and mutant hvKp strains with the expected phenotypes were obtained. These indicated that we established an efficient gene-editing method for genes on hvKp virulence plasmid, which could help us explore the functions of these genes and reveal the virulent mechanisms of hvKp.

本刊更多论文

高致病性肺炎克雷伯菌毒力质粒基因敲除的有效方法。

目前，高致病性肺炎克雷伯菌(hvKp)感染日益严重，其致毒机制尚不十分清楚。对hvKp毒力质粒上的基因进行有效的基因编辑可以帮助我们揭示相关的毒力机制。有一些报告侧重于上述方法，但有一定的局限性。在这项工作中，我们首先构建了基于pre112的重组自杀质粒，基于同源重组原理敲除或替换hvKp毒力质粒中的基因。结果表明，hvKp毒力质粒上的靶毒力基因iucA、iucB、iroB和rmpA2被无损伤敲除或被标记基因取代，获得了符合预期表型的hvKp突变株。这表明我们建立了一种高效的hvKp毒力质粒上基因的基因编辑方法，可以帮助我们探索这些基因的功能，揭示hvKp的毒力机制。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

New Microbiologica 生物-微生物学

CiteScore

2.20

自引率

5.60%

发文量

审稿时长

6-12 weeks

期刊介绍： The publication, diffusion and furtherance of research and study on all aspects of basic and clinical Microbiology and related fields are the chief aims of the journal.