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Received: 2019-06-22

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Journal of Zhejiang University SCIENCE B 2020 Vol.21 No.2 P.137-154

http://doi.org/10.1631/jzus.B1900440


Preliminary study on the role of novel LysR family gene kp05372 in Klebsiella pneumoniae of forest musk deer


Author(s):  Wei Yang, Wu-You Wang, Wei Zhao, Jian-Guo Cheng, Yin Wang, Xue-Ping Yao, Ze-Xiao Yang, Dong Yu, Yan Luo

Affiliation(s):  College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang 611130, China; more

Corresponding email(s):   lycjg@163.com

Key Words:  Moschus berezovskii, Klebsiella pneumoniae, LysR transcription factor, kp05372 gene, Biological characteristics, Upstream gene


Wei Yang, Wu-You Wang, Wei Zhao, Jian-Guo Cheng, Yin Wang, Xue-Ping Yao, Ze-Xiao Yang, Dong Yu, Yan Luo. Preliminary study on the role of novel LysR family gene kp05372 in Klebsiella pneumoniae of forest musk deer[J]. Journal of Zhejiang University Science B, 2020, 21(2): 137-154.

@article{title="Preliminary study on the role of novel LysR family gene kp05372 in Klebsiella pneumoniae of forest musk deer",
author="Wei Yang, Wu-You Wang, Wei Zhao, Jian-Guo Cheng, Yin Wang, Xue-Ping Yao, Ze-Xiao Yang, Dong Yu, Yan Luo",
journal="Journal of Zhejiang University Science B",
volume="21",
number="2",
pages="137-154",
year="2020",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1900440"
}

%0 Journal Article
%T Preliminary study on the role of novel LysR family gene kp05372 in Klebsiella pneumoniae of forest musk deer
%A Wei Yang
%A Wu-You Wang
%A Wei Zhao
%A Jian-Guo Cheng
%A Yin Wang
%A Xue-Ping Yao
%A Ze-Xiao Yang
%A Dong Yu
%A Yan Luo
%J Journal of Zhejiang University SCIENCE B
%V 21
%N 2
%P 137-154
%@ 1673-1581
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%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1900440

TY - JOUR
T1 - Preliminary study on the role of novel LysR family gene kp05372 in Klebsiella pneumoniae of forest musk deer
A1 - Wei Yang
A1 - Wu-You Wang
A1 - Wei Zhao
A1 - Jian-Guo Cheng
A1 - Yin Wang
A1 - Xue-Ping Yao
A1 - Ze-Xiao Yang
A1 - Dong Yu
A1 - Yan Luo
J0 - Journal of Zhejiang University Science B
VL - 21
IS - 2
SP - 137
EP - 154
%@ 1673-1581
Y1 - 2020
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1900440


Abstract: 
LysR-type transcriptional regulators are involved in the regulation of numerous cellular metabolic processes in Klebsiella pneumoniae, leading to severe infection. Earlier, we found a novel LysR family gene, named kp05372, in a strain of K. pneumoniae (designated GPKP) isolated from forest musk deer. To study the function of this gene in relation to the biological characteristics of GPKP, we used the suicide plasmid and conjugative transfer methods to construct deletion mutant strain GPKP-Δkp05372; moreover, we also constructed the GPKP-Δkp05372+ complemented strain. The role of this gene was determined by comparing the following characteristics of three strains: growth curves, biofilm formation, drug resistance, stress resistance, median lethal dose (LD50), organ colonization ability, and the histopathology of GPKP. Real-time polymerase chain reaction (RT-PCR) was used to test the expression level of seven genes upstream of kp05372. There was no significant difference in the growth rates when comparing the three bacterial strains, and no significant difference was recorded at different osmotic pressures, temperatures, salt contents, or hydrogen peroxide concentrations. The GPKP-Δkp05372 mutant formed a weak biofilm, and the other two strains formed medium biofilm. The drug resistance of the GPKP-Δkp05372 mutant toward cephalothin, cotrimoxazole, and polymyxin B was changed. The acid tolerance of the deletion strain was stronger than that of the other two strains. The LD50 values of the wild-type and complemented strains were 174-fold and 77-fold higher than that of the GPKP-Δkp05372 mutant, respectively. The colonization ability of the GPKP-Δkp05372 mutant in the heart, liver, spleen, kidney, and intestine was the weakest. The three strains caused different histopathological changes in the liver and lungs. In the GPKP-Δkp05372 mutant, the relative expression levels of kp05374 and kp05379 were increased to 1.32-fold and 1.42-fold, respectively, while the level of kp05378 was decreased by 42%. Overall, the deletion of kp05372 gene leads to changes in the following: drug resistance and acid tolerance; decreases in virulence, biofilm formation, and colonization ability of GPKP; and regulation of the upstream region of adjacent genes.

LysR家族新基因kp05372对林麝肺炎克雷伯氏菌的作用的初步研究

目的:通过kp05372基因的敲除来研究其对林麝肺炎克雷伯氏菌生物表型的影响.
创新点:解析了LysR家族新基因kp05372对林麝肺炎克雷伯氏菌生物表型的影响.
方法:通过基因敲除和回补技术构建了kp05372的基因缺失株和基因回补株,比较了kp05372基因缺失前后,林麝肺炎克雷伯氏菌在生长曲线、药敏试验、生物被膜、抗逆性、细菌半数致死量、定殖能力、病理形态学等生物学特性所表现出的差异.
结论:成功构建了kp05372基因的缺失株及回补株.kp05372基因的缺失导致林麝肺炎克雷伯氏菌耐药性和耐酸性发生了变化,毒力、生物膜和定殖能力降低,并且调控了上游临近区域的基因表达.

关键词:林麝;肺炎克雷伯氏菌;LysR转录因子;kp05372;生物表型;上游基因

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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