食源性单增李斯特菌inlA/inlB/inlC基因缺失株的构建及其生物学特性分析

doi:10.16431/j.cnki.1671-7236.2019.01.006

Abstract

Abstract:

In order to evaluate the effect of inlA/inlB/inlC genes on biological characteristics of Lm,the inlC gene deletion mutant was constructed by fusion PCR method,and then pKSV7-△inlC shuttle vector was constructed,which was transformed into competent cells Lm681-△inlAB.Homologous recombination was conducted using temperature (42℃)and chloramphenicol (10 μg/mL) resistance to achieve the inlC gene deletion strain,and then the homologous recombination was screened for identification and some biological characteristics were studied.PCR and sequencing results confirmed that Lm681-△inlABC was successfully obtained.The results showed that the growth characteristics of the deletion mutants were the same as the wild-type strain Lm681 and there was no significant difference.The hemolysis activity of the mutant strains were consistent with wild-type strain Lm681.In the mouse infection experiment,the mortality rate of wild-type strain Lm681,Lm681-△inlAB and Lm681-△inlABC were 80%(8/10),60%(6/10) and 40%(4/10),respectively.Results of mice virulence determination showed that the median lethal dose of wild-type strain,Lm681-△inlAB and Lm681-△inlABC were 4.36×10⁴,1.35×10⁶ and 2.95×10⁷ CFU,respectively.Furthermore,the colonization ability of the inlA/inlB/inlC genes deleted strain was extremely significantly lower than wild-type strain in liver,spleen and brain (P < 0.01).Collectively,these results indicated that inlA/inlB/inlC genes might play an important role in pathogenicity of Lm infection and provided scientific basis for the study on the mechanism in host cell invasion by Lm.

Key words: Listeria monocytogenes; internalin gene; gene mutant; biological characteristics

CLC Number:

S852.61

REN Jingjing, YANG Mingwei, CHEN Yunfei, YIN Hua, WANG Xinyu, LI Beibei, JIANG Jianjun, WANG Pengyan. Construction and Characterization of inlA/inlB/inlC Genes Deleted Mutant Strain of Food-borne Listeria monocytogenes[J]. , 2019, 46(1): 45-53.

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Construction and Characterization of inlA/inlB/inlC Genes Deleted Mutant Strain of Food-borne Listeria monocytogenes

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