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首页> 外文期刊>Infection and immunity >Characterization of vir-activated TnphoA gene fusions in Bordetella pertussis.
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Characterization of vir-activated TnphoA gene fusions in Bordetella pertussis.

机译：百日咳博德特氏菌中病毒激活的TnphoA基因融合体的表征。

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The expression of many of the known virulence determinants of Bordetella pertussis is coordinately regulated by the vir regulatory locus and reduced in response to environmental signals called modulators. We have previously identified eight TnphoA gene fusions in B. pertussis in which the expression of alkaline phosphatase was maximal in the absence of the modulators nicotinic acid and MgSO4. We have termed the genes identified by these fusions vir-activated genes. Here we report the characterization of these TnphoA mutant strains. Four fusion strains were defective in known virulence determinants. For one of these, fusion strain SK39, Southern blot hybridization demonstrated that TnphoA was inserted in the S1 subunit gene of pertussis toxin. Hemagglutination assays, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and immunoblots identified three fusions strains, SK16, SK75, and SK91, that were defective in filamentous hemagglutinin. Whereas all three filamentous hemagglutinin-defective mutants showed either normal or enhanced colonization, the pertussis toxin-defective mutant showed a marked defect in pulmonary persistence. Of the four other fusion strains, two were deficient in outer membrane proteins. One of these, strain SK8, was defective in a major outer membrane protein of 95 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This strain colonized mouse lungs less well and did not induce lymphocytosis after aerosol challenge. The other strain, SK34, was defective in four outer membrane proteins, three of which were detectable only on a Western blot with polyclonal sera against B. pertussis. Two of our gene fusion strains did not show any defect in identifiable vir-regulated proteins.
机译：百日咳博德特氏菌的许多已知毒力决定因素的表达受vir调节位点协调调节，并响应称为调节剂的环境信号而降低。我们先前已经确定了百日咳博德特氏菌中的八种TnphoA基因融合体，其中在缺乏调节剂烟酸和MgSO4的情况下碱性磷酸酶的表达最大。我们称这些融合蛋白为vir-activated基因。在这里，我们报告这些TnphoA突变株的表征。四种融合菌株在已知毒力决定因素方面存在缺陷。对于其中之一，融合菌株SK39，Southern印迹杂交证明TnphoA被插入百日咳毒素的S1亚基基因中。血凝测定，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和免疫印迹确定了丝状血凝素有缺陷的三种融合菌株SK16，SK75和SK91。尽管所有三个丝状血凝素缺陷型突变体均显示出正常或增强的定植，但百日咳毒素缺陷型突变体显示出肺持久性的明显缺陷。在其他四个融合菌株中，有两个在外膜蛋白方面缺乏。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测定，其中一个菌株SK8在95 kDa的主要外膜蛋白中存在缺陷。该菌株在小鼠肺部定植的效果较差，并且在气溶胶激发后未诱导淋巴细胞增多。另一株SK34在四个外膜蛋白中有缺陷，其中只有在针对百日咳博德特氏菌的多克隆血清的Western印迹中才能检测到其中的三个。我们的两个基因融合菌株在可识别的病毒调节蛋白中未显示任何缺陷。

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