食品科学2017，Vol.38Issue(10)：12-17,6.DOI:10.7506/spkx1002-6630-201710003
单增李斯特菌prfA基因缺失菌株的构建及其生物学特性鉴定
Construction and Biological Activity of a Strain of Listeria monocytogenes Deleted for Positive Regulatory Factor A Gene (prfA)
摘要
Abstract
Among the known Listeria strains,only Listeria monocytogenes (Lm) can cause human disease.The expression of most virulence genes is regulated fully or partially by positive regulatory factor A (PrfA) protein encoded by prfA gene.In this study,we adopted homologous recombination method to knock out the prfA gene of the wild strain EGDe,and investigated the biological characteristics of EGDe-△prfA by testing growth state,virulence gene expression and Caco-2 cell invasion.There was no difference in the growth status of EGDe-△prfA and EGDe as demonstrated by the growth curves.Real time quantitative PCR results showed that the expression levels ofplcA and plcB genes increased by 2.5 times and twice,respectively,while the expression levels of inlA,inlB,inlC,actA and vip declined.The expression level ofprfA gene was nearly zero.The invasion of Caco-2 cells by EGDe-△prfA was one fifth as high as that by the wild strain.The gene-deleted strain can provide an important tool for studying the pathogenic mechanism of the foodbome pathogen Listeria monocytogenes.关键词
单核细胞增生性李斯特菌/prfA/基因敲除/生物学特性Key words
Listeria monocytogenes/prfA/gene knock-out/biological characteristics分类
轻工纺织引用本文复制引用
郭亮,陈国薇,谢曼曼,刘武康,丁承超,王淑娟,罗勤,刘箐..单增李斯特菌prfA基因缺失菌株的构建及其生物学特性鉴定[J].食品科学,2017,38(10):12-17,6.基金项目
国家自然科学基金面上项目(31371776) （31371776）
上海市科委“科技创新行动计划”长三角科技联合攻关领域项目(15395810900) （15395810900）
上海理工大学研究生创新基金资助项目 （）
上海理工大学微创励志创新基金项目 （）
