Pseudomonas fluorescens adhesion and transport through porous media are affected by lipopolysaccharide composition
Pseudomonas fluorescens adhesion and transport through porous media are affected by lipopolysaccharide composition
复制标题
DOI：
10.1128/aem.62.1.100-104.1996
复制
发表时间：
1996-01-01
影响因子：
4.4
通讯作者：
Fletcher, M
中科院分区：
文献类型：
--
作者：
Williams, V;Fletcher, M
这项工作的目的是（i）使用转座子诱变产生的突变体的荧光假单胞菌的粘附能力和运输通过多孔介质的改变和（ii），以确定在表面特性的改变，负责的变化，附件。用TnphoA产生荧光假单胞菌的突变体，这使得能够鉴定表面蛋白改变的突变体。通过在疏水聚苯乙烯和水溶性聚苯乙烯上的附着测定筛选转座子突变体的粘附能力改变。获得了四种TnphoA突变体，其对疏水表面的粘附增加，对水溶性表面的粘附减少。通过使每种突变体和亲本的悬浮液通过含有石英砂的柱并确定保留在柱中的细胞数来评价菌株通过多孔介质的运输。突变体都表现出增加的粘附和保留在列中。Southern分析证明了两种类型的突变体具有不同的转座子插入位点。聚丙烯酰胺凝胶电泳的菌株表明，O抗原的脂多糖是减弱或不存在。缺乏这种多糖，以及随之而来的脂多糖的脂质部分的暴露增加，可能是负责增加粘附到疏水基质和保留在砂柱。本工作结合了以前对荧光假单胞菌附着的研究，证明了不止一种类型的聚合物可以介导该生物体对非生物表面的粘附。
The objectives of this work were (i) to use transposon mutagenesis to produce mutants of Pseudomonas fluorescens that were altered in adhesion ability and transport through porous media and (ii) to identify the alterations in surface characteristics that were responsible for the changes in attachment. Mutants of P. fluorescens were generated with TnphoA, which enabled identification of mutants that were altered in surface proteins, Transposon mutants were screened for alterations in adhesion ability by attachment assays on hydrophobic polystyrene and water-wettable polystyrene. Four TnphoA mutants with increased adhesion to the hydrophobic surface and decreased adhesion to the water-wettable surface were obtained. Transport of the strains through porous media was evaluated by passing suspensions of each mutant and the parent through columns containing quartz sand and determining the number of cells retained in the columns. The mutants all demonstrated increased adhesion and retention in the columns. Southern analysis demonstrated two types of mutants with separate transposon insertion sites. Polyacrylamide gel electrophoresis of the strains demonstrated that the O antigen on the lipopolysaccharide was either attenuated or absent. Lack of this polysaccharide, and the consequent increased exposure of the lipid moiety of the lipopolysaccharide, is probably responsible for the increase in adhesion to the hydrophobic substrata and retention in the sand column. This work combined viith previous studies of attachment of P, fluorescens demonstrates that more than one type of polymer can mediate the adhesion of this organism to nonbiological surfaces.