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首页> 外文期刊>Infection and immunity >Salmonella enterica Serovar Typhi Uses Type IVB Pili To Enter Human Intestinal Epithelial Cells
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Salmonella enterica Serovar Typhi Uses Type IVB Pili To Enter Human Intestinal Epithelial Cells

机译：肠炎沙门氏菌伤寒沙门氏菌使用IVB型菌毛进入人肠上皮细胞

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DNA sequencing upstream of the Salmonella entericaserovar Typhi pilV and rci genes previously identified in the ca. 118-kb major pathogenicity island (X.-L. Zhang, C. Morris, and J. Hackett, Gene 202:139–146, 1997) identified a further 10 pil genes apparently forming a piloperon. The product of the pilS gene, prePilS protein (a putative type IVB structural prepilin) was purified, and an anti-prePilS antiserum was raised in mice. Mutants of serovar Typhi either lacking the whole pil operon or with an insertion mutation in the pilS gene were constructed, as was a strain in which the pilN to pilV genes were driven by the tac promoter. The pil + strains synthesized type IVB pili, as judged by (i) visualization in the electron microscope of thin pili in culture supernatants of one such strain and (ii) the presence of PilS protein (smaller than the prePilS protein by removal of the leader peptide) on immunoblotting of material pelleted by high-speed centrifugation of either the culture supernatant or sonicates of pil + strains. Controlpil mutants did not express the PilS protein. ApilS mutant of serovar Typhi entered human intestinal INT407 cells in culture to levels only 5 to 25% of those of the wild-type strain, and serovar Typhi entry was strongly inhibited by soluble prePilS protein (50% inhibition of entry at 1.4 μM prePilS).
机译：沙门氏菌沙门氏菌伤寒沙门氏菌pilV和rci基因上游的DNA测序先前在ca. 118 kb的主要致病岛（X.-L. Zhang，C。Morris和J. Hackett，Gene 202：139-146，1997）确定了另外10个pil基因，它们显然形成了菌毛子。纯化pilS基因的产物prePilS蛋白（推定的IVB型结构prepilin），并在小鼠中产生抗prePilS抗血清。构建了缺少完整pil操纵子或在pilS基因中具有插入突变的血清型Typhi突变体，以及由tac启动子驱动pilN到pilV基因的菌株。 pil +菌株合成了IVB型菌毛，这是通过以下方式判断的：（i）在一种此类菌株的培养上清液中的细菌毛的电子显微镜中观察，以及（ii）PilS蛋白的存在（通过去除前导区而小于prePilS蛋白）通过对pil +菌株的培养上清液或超声波进行高速离心沉淀的物质进行免疫印迹分析。 Controlpil突变体不表达PilS蛋白。血清型伤寒菌的ApilS突变体进入培养的人肠道INT407细胞的水平仅为野生型菌株的5％至25 ％，并且可溶性prePilS蛋白强烈抑制了血清型伤寒菌的进入（1.4μM时50％的进入抑制）准备）。

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