LIPI-4 EⅡC调控单增李斯特菌关键毒力因子转录的研究

doi:10.16431/j.cnki.1671-7236.2023.04.006

Abstract

Abstract: 【Objective】 This study was aimed to investigate the effect of membrane permease ⅡC(EⅡC) gene in Listeria monocytogenes (LM) pathogenicity island 4 (LIPI-4) on the key virulence factor genes expression.【Method】 The EⅡC gene deletion strain LM928ΔEⅡC and complementary strain CLM928ΔEⅡC of isolate LM928 were constructed.The growth curve of each strain and the fermentation of different sugars were determined.The adhesion,invasion and intracellular proliferation of each strain to human brain microvascular endothelial cells (HCMEC/D3) were determined.The transcript levels of virulence genes of each strain were detected using Real-time quantitative PCR under BHI medium and HCMEC/D3.【Result】 LM928 EⅡC gene deletion strain LM928ΔEⅡC and complementary strain CLM928ΔEⅡC were successfully constructed.Compared with the LM928,the LM928ΔEⅡC strain showed consistent growth curves and fermentability to glucose,cellobiose,fructose,rhamnose,esculin and salicin.The invasive ability of LM928ΔEⅡC decreased extremely significantly (P＜0.01), but the adhesion ability was not change significantly (P＞0.05).The amount of intracellular proliferation of LM928ΔEⅡC was significantly lower than that of LM928 and CLM928ΔEⅡC from 2 to 4 h post-infection (P＜0.05),but it was significantly higher than that of LM928 and CLM928ΔEⅡC from 8 to 12 h post-infection (P＜0.05).Compared with LM928 and CLM928ΔEⅡC,in BHI medium,the transcriptional levels of hly,prfA,actA,plcB, inlA,inlB,inlC and mpl genes of LM928ΔEⅡC were extremely significantly down-regulated (P＜0.01),the transcriptional level of plcA gene was extremely significantly up-regulated (P＜0.01).While in HCMEC/D3 cell,the transcriptional levels of hly,prfA,inlB,inlC and mpl genes of LM928ΔEⅡC were significant or extremely significantly up-regulated (P<0.05 or P<0.01),the transcriptional levels of plcA and inlA genes were extremely significant down-regulated (P＜0.01),and there was no significant difference between the transcriptional levels of actA and plcB genes.【Conclusion】 EⅡC in LIPI-4 did not affect the growth and partial sugar fermentation of LM928,could regulate important invasion-related genes and virulence-related genes in LM,which laid a foundation for further exploration of the role of EⅡC in LM and LIPI-4.

Key words: Listeria monocytogenes; pathogenicity island 4; membrane permease ⅡC; virulence gene

CLC Number:

S852.61

LIU Caixia, SHI Weidi, KOU Lijun, MA Xun, WANG Jing, GAO Shengjie, LYU Shuangfei, CHEN Xuke, ZENG Dongdong, KONG Cuilian, REN Huijie. Regulation of Transcription of Key Virulence Factors of Listeria monocytogenes by LIPI-4 EⅡC[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1340-1351.

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Regulation of Transcription of Key Virulence Factors of Listeria monocytogenes by LIPI-4 EⅡC

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