REGULATION BY A NOVEL PROTEIN OF THE BIMODAL DISTRIBUTION OF LIPOPOLYSACCHARIDE IN THE OUTER-MEMBRANE OF ESCHERICHIA-COLI
REGULATION BY A NOVEL PROTEIN OF THE BIMODAL DISTRIBUTION OF LIPOPOLYSACCHARIDE IN THE OUTER-MEMBRANE OF ESCHERICHIA-COLI
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DOI：
10.1128/jb.173.18.5699-5704.1991
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发表时间：
1991-09-01
影响因子：
3.2
通讯作者：
HULL, SI
中科院分区：
文献类型：
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作者：
BATCHELOR, RA;HARAGUCHI, GE;HULL, SI
我们报告的O 75脂多糖的rfb基因簇的大肠杆菌泌尿道分离株的克隆和表征。 缺失克隆确定了表达O 75抗原的最小DNA区域。coli宿主菌株上的12.4-kb插入片段。 然而，E.十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和银染检测表明，表达该区域的大肠杆菌菌株不产生聚合的O链。 一个稍大的13.4kb DNA克隆在E. coliSempty-set 874，但发现其在表面膜上的分布是异常的。 正常野生型E.与沙门氏菌属一样，具有脂多糖在表面上的双峰分布，其被看作是附着于脂质A-核心结构的长和短O链的丰度。 我们发现，在一个区域相邻的克隆rfb区域，并在相对侧从推定的聚合酶（rfc）编码，一种新的蛋白质的35.5 kDa的表达从1.75 kb的DNA片段。 这种蛋白质被证明是反式互补的E。大肠杆菌菌株携带质粒表达异常，不受管制的脂多糖。 这些互补菌株的表达呈双峰分布。 编码该蛋白的基因的突变破坏了其调节O链分布的能力。 我们建议将此调节基因称为rol，即O长度调节基因。
We report on the cloning and characterization of the rfb gene cluster of the O75 lipopolysaccharide from a urinary tract isolate of Escherichia coli. Deletion cloning defined the minimum region of DNA that expressed the O75 antigen in E. coli host strains to be on a 12.4-kb insert. However, the E. coli strain expressing this region did not produce a polymerized O chain as detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining. A slightly larger DNA clone of 13.4 kb produced a polymerized O chain in E. coli S empty-set 874 but was found to be abnormal in its distribution over the surface membrane. Normal wild-type E. coli, as with Salmonella spp., has a bimodal distribution of the lipopolysaccharide on the surface which is seen as an abundance of long and short O chains attached to the lipid A-core structure. We found in a region adjacent to the cloned rfb region, and on the opposite side from where the putative polymerase (rfc) is encoded, a novel protein of 35.5 kDa expressed from a 1.75-kb DNA fragment. This protein was shown to complement in trans the E. coli strains carrying plasmids that expressed abnormal, unregulated lipopolysaccharides. The expression of these complemented strains was bimodal in distribution. Mutation of the gene encoding this protein destroyed its ability to regulate O-chain distribution. We propose to call this regulator gene rol, for regulator of O length.