Preliminary study on the role of novel LysR family gene kp05372 in Klebsiella pneumoniae of forest musk deer
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 de...
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Published in | Journal of Zhejiang University. B. Science Vol. 21; no. 2; pp. 137 - 154 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Hangzhou
Zhejiang University Press
01.02.2020
Springer Nature B.V |
Subjects | |
Online Access | Get full text |
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Summary: | 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-Δ
kp05372
; moreover, we also constructed the GPKP-Δ
kp05372
+
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 (LD
50
), 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-Δ
kp05372
mutant formed a weak biofilm, and the other two strains formed medium biofilm. The drug resistance of the GPKP-Δ
kp05372
mutant toward cephalothin, cotrimoxazole, and polymyxin B was changed. The acid tolerance of the deletion strain was strongerthan that of the othertwo strains. The LD50 values of the wild-type and complemented strains were 174-fold and 77-fold higher than that of the GPKP-Δ
kp05372
mutant, respectively. The colonization ability of the GPKP-Δ
kp05372
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-Δ
kp05372
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. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Corresponding Author The two authors contributed equally to this work |
ISSN: | 1673-1581 1862-1783 |
DOI: | 10.1631/jzus.B1900440 |