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
发布时间:2018-10-11 10:38
【摘要】:单核细胞增生性李斯特菌(Listeria monocytogenes,Lm)是一种风险较高的食源性致病菌,其绝大多数毒力基因的表达均受到由prfA基因编码的PrfA(positive regulatory factorA)蛋白全部或部分调控。使用同源重组的方法敲除Lm野生菌株EGDe的prfA基因,并通过测定生长曲线、毒力基因表达和侵袭Caco-2细胞能力等探讨单缺失菌株EGDe-ΔprfA生物学特性。通过测定生长曲线显示prfA敲除株和野生型EGDe二者生长状态无差异;运用实时定量荧光聚合酶链式反应检测EGDe-ΔprfA的主要毒力基因表达,结果显示plcA、plcB毒力基因的表达量分别上升至原来的2.5倍和2倍,inlA、inlB、inlC、actA、vip等均呈下降趋势,prfA基因表达量趋向于零;侵袭Caco-2细胞结果显示EGDe-ΔprfA侵袭数为野生株的1/5。该基因缺失菌株的构建及生物学特性研究对食源性致病菌EGDe致病机理研究具有重要意义并且提供了重要材料。
[Abstract]:Listeria monocytogenes (Listeria monocytogenes,Lm) is a kind of foodborne pathogenic bacteria with high risk. The expression of most virulence genes of Listeria monocytogenes (Listeria monocytogenes,Lm) is regulated in whole or in part by the PrfA (positive regulatory factorA) protein encoded by prfA gene. Homologous recombination was used to knockout the prfA gene of Lm wild strain EGDe, and the biological characteristics of single deletion strain EGDe- 螖 prfA were studied by measuring growth curve, virulence gene expression and ability of invading Caco-2 cells. The growth curve showed that there was no difference between prfA knockout and wild-type EGDe, and the expression of main virulence genes of EGDe- 螖 prfA was detected by real-time quantitative fluorescence polymerase chain reaction. The results showed that the expression of virulence gene of plcA,plcB increased to 2.5 times and 2 times of the original, inlA,inlB,inlC,actA,vip and so on showed a downward trend, prfA gene expression tended to zero, and invasion of Caco-2 cells showed that the number of EGDe- 螖 prfA invasion was 1 / 5 of that of the wild strain. The construction and biological characteristics of this gene deletion strain are of great significance for the study of pathogenic mechanism of foodborne pathogenic bacteria EGDe and provide important materials.
【作者单位】: 上海理工大学医疗器械与食品学院;
【基金】:国家自然科学基金面上项目(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) 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
【分类号】:TS201.3
[Abstract]:Listeria monocytogenes (Listeria monocytogenes,Lm) is a kind of foodborne pathogenic bacteria with high risk. The expression of most virulence genes of Listeria monocytogenes (Listeria monocytogenes,Lm) is regulated in whole or in part by the PrfA (positive regulatory factorA) protein encoded by prfA gene. Homologous recombination was used to knockout the prfA gene of Lm wild strain EGDe, and the biological characteristics of single deletion strain EGDe- 螖 prfA were studied by measuring growth curve, virulence gene expression and ability of invading Caco-2 cells. The growth curve showed that there was no difference between prfA knockout and wild-type EGDe, and the expression of main virulence genes of EGDe- 螖 prfA was detected by real-time quantitative fluorescence polymerase chain reaction. The results showed that the expression of virulence gene of plcA,plcB increased to 2.5 times and 2 times of the original, inlA,inlB,inlC,actA,vip and so on showed a downward trend, prfA gene expression tended to zero, and invasion of Caco-2 cells showed that the number of EGDe- 螖 prfA invasion was 1 / 5 of that of the wild strain. The construction and biological characteristics of this gene deletion strain are of great significance for the study of pathogenic mechanism of foodborne pathogenic bacteria EGDe and provide important materials.
【作者单位】: 上海理工大学医疗器械与食品学院;
【基金】:国家自然科学基金面上项目(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) 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
【分类号】:TS201.3
【相似文献】
相关期刊论文 前10条
1 徐来杰,王月娥;稻纵卷叶螟绒茧蜂生物学特性初步研究[J];植保技术与推广;1999年06期
2 顾峰;张贵军;李宝贵;范志明;曹淑兰;;吉林省小肠结肠炎耶尔森氏菌生物学特性研究[J];中国卫生工程学;2006年05期
3 崔宗均,路鹏,于会泳,张晋丰;高温菌生物学特性的应用[J];微生物学杂志;2003年04期
4 徐承水;磷脂的生物学特性及保健功能[J];临沂师范学院学报;2000年03期
5 张大治,于有志,任国栋;心形刺甲的生物学特性[J];宁夏农学院学报;2000年04期
6 易力;汪洋;范春永;李营营;;鸽源乳酸杆菌的分离鉴定与生物学特性研究[J];养禽与禽病防治;2008年08期
7 严林;杜艳丽;郭蕊;郝金娥;刘长仲;;灰钝额斑螟的生物学特性[J];应用昆虫学报;2011年06期
8 毛本勇;突缘拟凹背蝗生物学特性初步研究[J];大理师专学报(综合版);1998年02期
9 杨大荣,沈发荣,杨跃雄,梁醒财,董大志,春生,鲁自,斯那都吉;云南蝠蛾属一新种及生物学特性研究[J];昆虫学报;1991年02期
10 欧汉标,莫冠英;雷州半岛臭,
本文编号:2263890
本文链接:https://www.wllwen.com/kejilunwen/jiyingongcheng/2263890.html
最近更新
教材专著