Pathogenesis and molecular genetics of expression of iron acquisition systems in Vibro parahaemolyticus

副溶血弧菌铁获取系统表达的发病机制和分子遗传学

• 批准号: 07670312
• 负责人: YAMAMOTO Shigeo
• 金额: $ 1.41万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670312/
• 关键词: Vibrio parahaemolyticus; Iron acquisition; Iron-repressible gene; Repressor protein; Exocellular protease; Molecular cloning; Pathogenic factor; Mutant; シデロフォア; ヘム鉄; Furレギュロン; 外膜レセプター

1. The 83-kDa iron-repressible outer membrane protein which binds hemin was purified and the antiserum against this protein was raised in rabbits. Immunoblot analysis using this antiserum indicated that the molecular mass and antigenic properties of the protein were highly conserved among this species.2. A total of 109 V.parahaemolyticus strains isolated from different sources were examined for their activity to produced vibrioferrin in spend cultures, which was determined by a new developed HPLC method. Vibrioferrin was detected in most of the strains tested (95.2%). The mean value <plus-minus> SD of vibrioferrin in the clinical isolates (n=44) was 742.7 <plus-minus> 46.7 muM in sup./OD660, signifcantly different with p<0.01 from the mean values in food isolate (386.5 <plus-minus> 43.6, n=35) and environmental isolates (378,8 <plus-minus> 37.2, n=25). These results suggest that the higher productivity of clinical isolates may afford a significant selective advantage for growth in cond … More itions of iron-limitation such as exist in the in3. Manganese resistant mutants of V.parahaemolyticus WP1 were isolated and characterized. The mutants expressed the 78- and 83-kDa outer membrane proteins independently of iron, suggesting the existence of an Fur-regulon. The fur gene, encoding an iron-regulatory protein has been cloned and sequenced. Its product showed a high degree of homology to known Fur sequence.4. Exocellular protease activity of V.parahaemolyticus strains was significantly enhanced when they were grown under iron-limited conditions. Some manganese resistant mutants exhibited higher protease activity than the wild type strain WP1 even when grown under iron sufficient conditions, suggesting that expression of the gene encoding a protease (s) may be regulated by the Fur.Further study is required to clarify as to what kind of protease gene is regulated by Fur, since the presence of four proteases has been reported for this species.Many clones which may have genes regulated by Fur were isolated by Fur titration assay method. Further nucleotide sequence analysis of these genes may lead to identification of iron-responsive genes and to cloning of the whole genes responsible. Less

1.纯化了结合氯化血红素的83-kDa铁抑制性外膜蛋白，并在兔中产生了针对该蛋白的抗血清。利用该抗血清进行免疫印迹分析，结果表明该蛋白的分子量和抗原特性在该物种中高度保守.采用高效液相色谱法测定了109株不同来源的副溶血性弧菌在培养液中产生弧菌铁蛋白的活性。在大多数测试菌株中检测到弧菌铁蛋白（95.2%）。临床<plus-minus>分离株（n=44）中弧菌铁蛋白的平均值SD为742.7 <plus-minus>± 46.7 μ M，OD_（660），与食品分离株（386.5 <plus-minus>± 43.6，n=35）和环境分离株（378，8 ± <plus-minus>37.2，n=25）的平均值有显著性差异（p&lt;0.01）。这些结果表明，较高的生产力的临床分离株可能提供了一个显着的选择性优势，生长在条件 ...更多信息 铁限制的情况如存在于IN 3中。分离并鉴定了副溶血性弧菌WP 1的锰抗性突变体。突变体表达的78-和83-kDa的外膜蛋白独立的铁，这表明存在的毛皮调节子。编码铁调节蛋白的fur基因已被克隆和测序。其产物与已知的Fur序列具有高度的同源性.当副溶血性弧菌菌株在铁限制条件下生长时，其胞外蛋白酶活性显著增强。一些锰抗性突变株即使在铁充足的条件下生长也表现出比野生型菌株WP 1更高的蛋白酶活性，这表明编码蛋白酶的基因的表达可能受到Fur的调节。由于已报道该物种存在四种蛋白酶，因此通过Fur滴定法分离了许多可能具有受Fur调控的基因的克隆。这些基因的进一步核苷酸序列分析可能导致鉴定铁响应基因和克隆的整个基因负责。少

项目成果

山本重雄: "病原菌の鉄獲得機構" 日本細菌学雑誌. 51. 523-547 (1996)

Shigeo Yamamoto：“病原菌的铁获取机制”日本细菌学杂志 51. 523-547 (1996)。

Shigeo Yamamoto: "Utilization of hemin and hemoglobin as iron sources by Vibrio parahaemolyticus and identification of an iron-repressible hemin-binding protein." FEMS Microbiology Letters.128. 195-200 (1995)

Shigeo Yamamoto：“副溶血弧菌利用血红素和血红蛋白作为铁源，并鉴定了铁抑制型血红素结合蛋白。”

Shigeo Yamamoto: "Utilization of hemin and hemoglobin as iron sources by Vibrio parahaemolyticus and identification of an iro-repressible hemin-binding protein" FEMS Microbiology Letters. 128. 195-200 (1995)

Shigeo Yamamoto：“副溶血弧菌利用血红素和血红蛋白作为铁源以及铁抑制血红素结合蛋白的鉴定”FEMS 微生物学快报。

Shigeo Yamamoto: "Siderophores-Iron transporting agent produced by microorganisms." Pharmacia. 31. 588-592 (1995)

Shigeo Yamamoto：“铁载体——微生物产生的铁转运剂。”

Nunirul Alam: "Expression of virulence-related properties by, and intestinal adhesiveness of, Vibrio mimicus strains isolated from aquatic environment." Applied Environmental Microbiology. 62. 3871-3874 (1996)

Nunirul Alam：“从水生环境中分离出的拟态弧菌菌株的毒力相关特性及其肠道粘附性的表达。”