Identification of Listeria monocytogenes determinants required for biofilm formation.
Identification of Listeria monocytogenes determinants required for biofilm formation.
复制标题
DOI：
10.1371/journal.pone.0113696
复制
发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Higgins DE
中科院分区：
文献类型：
--
作者：
Alonso AN;Perry KJ;Regeimbal JM;Regan PM;Higgins DE
单核细胞增生李斯特菌是一种革兰氏阳性的食源性致病菌。L.美国食品和药物管理局（FDA）认为单核细胞增多症是一种潜在的公共卫生风险，因为这种细菌很容易污染即食（RTE）食品，并导致侵入性的危及生命的疾病（霍乱）。细菌可以在多个表面上粘附和生长，并在食品加工厂的生物膜内持续存在，从而对消毒剂和其他抗菌剂产生抗性。虽然全基因组测序已经导致在许多细菌物种中鉴定生物膜合成基因簇，但生物信息学尚未鉴定L.单核细胞增生基因组。为了鉴定L.为了抑制单核细胞增多症生物膜形成，我们使用最近构建的Himar 1 mariner转座子进行了转座子诱变文库筛选。在L.在96孔聚氯乙烯（PVC）微量滴定板中筛选单核细胞增生菌株10403S的生物膜形成，其中鉴定出70个Himar 1插入突变体，其产生显著较少的生物膜。分离的突变体内的转座子插入位点的DNA测序揭示转座子插入38个不同的遗传位点内。在先前已知的生物膜形成的初始阶段所需的几个鞭毛运动性基因中携带插入物的突变体的鉴定验证了诱变筛选鉴定L.单核细胞增多症生物膜缺陷突变体。选择两个新鉴定的遗传位点dltABCD和phoPR进行缺失分析，ΔdltABCD和ΔphoPR细菌菌株在PVC微量滴定板测定中均显示生物膜形成缺陷，证实这些位点有助于L.单核细胞增多症
Listeria monocytogenes is a Gram-positive, food-borne pathogen of humans and animals. L. monocytogenes is considered to be a potential public health risk by the U.S. Food and Drug Administration (FDA), as this bacterium can easily contaminate ready-to-eat (RTE) foods and cause an invasive, life-threatening disease (listeriosis). Bacteria can adhere and grow on multiple surfaces and persist within biofilms in food processing plants, providing resistance to sanitizers and other antimicrobial agents. While whole genome sequencing has led to the identification of biofilm synthesis gene clusters in many bacterial species, bioinformatics has not identified the biofilm synthesis genes within the L. monocytogenes genome. To identify genes necessary for L. monocytogenes biofilm formation, we performed a transposon mutagenesis library screen using a recently constructed Himar1 mariner transposon. Approximately 10,000 transposon mutants within L. monocytogenes strain 10403S were screened for biofilm formation in 96-well polyvinyl chloride (PVC) microtiter plates with 70 Himar1 insertion mutants identified that produced significantly less biofilms. DNA sequencing of the transposon insertion sites within the isolated mutants revealed transposon insertions within 38 distinct genetic loci. The identification of mutants bearing insertions within several flagellar motility genes previously known to be required for the initial stages of biofilm formation validated the ability of the mutagenesis screen to identify L. monocytogenes biofilm-defective mutants. Two newly identified genetic loci, dltABCD and phoPR, were selected for deletion analysis and both ΔdltABCD and ΔphoPR bacterial strains displayed biofilm formation defects in the PVC microtiter plate assay, confirming these loci contribute to biofilm formation by L. monocytogenes.
登录
查看更多内容
血素素在单核细胞增生李斯特菌的细胞内生长中的作用。
DOI：
10.1084/jem.167.4.1459
发表时间：
1988-04-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Portnoy DA;Jacks PS;Hinrichs DJ
通讯作者：
Hinrichs DJ
影响因子：
6.7
作者：
Saar-Dover R;Bitler A;Nezer R;Shmuel-Galia L;Firon A;Shimoni E;Trieu-Cuot P;Shai Y
通讯作者：
Shai Y
影响因子：
3.2
作者：
Perry, Kyle J.;Higgins, Darren E.
通讯作者：
Higgins, Darren E.
影响因子：
3.2
作者：
Kocan, M;Schaffer, S;Bott, M
通讯作者：
Bott, M
影响因子：
3.1
作者：
GAILLARD, JL;BERCHE, P;SANSONETTI, P
通讯作者：
SANSONETTI, P