The First Dongguan Affiliated Hospital, Guangdong Medical University No.42 Jiaoping Avenue, Tangxia Town, Dongguan City, 523000, Guangdong Province, China.
Department of Clinical Laboratory, Affiliated Hospital of Guangdong Medical University, No. 57, South Renmin Avenue, Xiashan District, Zhanjiang City, 524000 Guangdong Province, China.
New Microbiol. 2023 May;46(2):186-195.
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.
目前，高毒力肺炎克雷伯菌（hvKp）的感染日趋严重，但其毒力机制尚不清楚。一种有效的 hvKp 毒力质粒上基因的编辑方法可以帮助我们揭示相关的毒力机制。目前已有一些关于上述方法的报道，但存在一定的局限性。在本工作中，我们首先构建了基于 pRE112 的重组自杀质粒，根据同源重组原理对 hvKp 毒力质粒上的基因进行敲除或替换。结果表明，目标毒力基因 iucA、iucB、iroB 和 rmpA2 在 hvKp 毒力质粒上被无痕敲除或被标记基因替换，并获得了具有预期表型的突变 hvKp 菌株。这表明我们建立了一种高效的 hvKp 毒力质粒上基因编辑方法，可以帮助我们研究这些基因的功能，揭示 hvKp 的毒力机制。
