CHARACTERIZATION OF TRANSPOSON MUTANTS OF BIOFILM-PRODUCING STAPHYLOCOCCUS-EPIDERMIDIS IMPAIRED IN THE ACCUMULATIVE PHASE OF BIOFILM PRODUCTION - GENETIC IDENTIFICATION OF A HEXOSAMINE-CONTAINING POLYSACCHARIDE INTERCELLULAR ADHESIN
CHARACTERIZATION OF TRANSPOSON MUTANTS OF BIOFILM-PRODUCING STAPHYLOCOCCUS-EPIDERMIDIS IMPAIRED IN THE ACCUMULATIVE PHASE OF BIOFILM PRODUCTION - GENETIC IDENTIFICATION OF A HEXOSAMINE-CONTAINING POLYSACCHARIDE INTERCELLULAR ADHESIN
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DOI：
10.1128/iai.62.8.3244-3253.1994
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发表时间：
1994-08-01
影响因子：
3.1
通讯作者：
LAUFS, R
中科院分区：
文献类型：
--
作者：
MACK, D;NEDELMANN, M;LAUFS, R
首先附着在聚合物表面，然后在多层细胞团中积累，导致表皮葡萄球菌生物膜的产生，这种生物膜被认为有助于生物材料相关感染的毒力。我们分离到生物膜产生型表皮葡萄球菌13-1的Tn917转座子突变株，其生物膜完全阴性。在脉冲场凝胶电泳法中，未发现突变体的明显缺失。突变体M10和M11的Tn917插入位于不同的EcoRI片段上，但位于相同的60kb SmaI和17kb BamHI染色体片段上。噬菌体转导显示突变株M10和M11的转座子插入与改变的表型相关联，而其他几个突变株则明显表现为自发变异。在聚苯乙烯小球的初步附着试验中，没有检测到任何突变体与野生型之间的显著差异。细胞聚集作为细胞间黏附的指示，这是在多层细胞团中积累的先决条件，在任何突变体中都没有检测到。这些结果表明，突变体在生物被膜产生的积累阶段受到了损害。突变体M10和M11不产生可检测到的量的特定多糖抗原(II.Mack、N.Siemssen和R.Lauff感染。伊蒙。60：2048-2057,1992)，而自发变异体产生的抗原量大幅减少。由于同基因生物膜阴性转导子1457-M11制备的抗原中几乎没有检测到己糖胺，因此凝胶过滤富集产生生物膜的表皮葡萄球菌1457抗原的主要特异组分为己糖胺，这是该抗原与其他表皮葡萄球菌多糖成分的区别。我们的结果为该抗原在表皮葡萄球菌生物膜形成的累积阶段通过介导细胞间黏附而发挥作用提供了直接的遗传学证据。
The primary attachment to polymer surfaces followed by accumulation in multilayered cell clusters leads to production of Staphylococcus epidermis biofilms, which are thought to contribute to virulence in biomaterial-related infections. We isolated Tn917 transposon mutants of biofilm-producing S. epidermidis 13-1, which were completely biofilm negative. In pulsed-field gel electrophoresis no obvious deletions of the mutants were noted. The Tn917 insertions of mutants M10 and M11 were located on different EcoRI fragments but on identical 60-kb SmaI and 17-kb BamHI chromosomal fragments. Linkage of transposon insertions of mutants M10 and M11 with the altered phenotype was demonstrated by phage transduction, whereas the several other mutants apparently represented spontaneous variants. In a primary attachment assay with polystyrene spheres, no significant difference between any of the mutants and the wild type could be detected. Cell clustering as an indication of intercellular adhesion, which is a prerequisite for accumulation in multilayered cell clusters, was not detected with any mutant. These results demonstrate that the mutants were impaired in the accumulative phase of biofilm production. Mutants M10 and M11 did not produce detectable amounts of a specific polysaccharide antigen (Ii. Mack, N. Siemssen, and R. Laufs, Infect. Immun. 60:2048-2057, 1992), whereas substantially reduced amounts of antigen were produced by the spontaneous variants. Hexosamine was determined as the major specific component of the antigen enriched by gel filtration of biofilm-producing S. epidermidis 1457 because almost no hexosamine was detected in material prepared from the isogenic biofilm-negative transductant 1457-M11, which differentiates the antigen from other S. epidermidis polysaccharide components. Our results provide direct genetic evidence for a function of the antigen in the accumulative phase of biofilm production by S. epidermidis by mediating intercellular adhesion.