Identification and characterization of the TonB region and its role in transferrin-mediated iron acquisition in Haemophilus parasuis
Identification and characterization of the TonB region and its role in transferrin-mediated iron acquisition in Haemophilus parasuis
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DOI：
10.1016/j.femsim.2005.02.008
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发表时间：
2005-07-01
影响因子：
--
通讯作者：
Rodríguez-Ferri, EF
中科院分区：
文献类型：
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作者：
del Río, ML;Gutiérrez-Martín, CB;Rodríguez-Ferri, EF
副猪嗜血杆菌是引起格拉斯氏病的病原，给养猪业造成了巨大的经济损失。这项研究的目的是对包括TonB的TonB区域进行鉴定、测序和分子特征分析。副猪嗜血杆菌的exbbd和tbpBA基因。此外，还产生了两种融合蛋白。其中一个(pGEX-6P-1-GST-TbpB)含有副猪嗜血杆菌TbpB蛋白的前501个氨基酸，第二个(pBAD-Thio-TbpB-V5-His)含有副猪嗜血杆菌TbpB N端区的前102个氨基酸。制备了14株分泌抗两种重组TbpB融合蛋白的单抗的杂交瘤细胞。此外，为了评价在铁限制条件下副猪嗜血杆菌ExbB、TbpB和TbpA蛋白的表达是否上调，制备了抗副猪嗜血杆菌TbpB-His融合蛋白的兔多克隆抗体。用兔抗7型胸膜肺炎放线杆菌ExbB和TbpA蛋白的多克隆抗体检测副猪嗜血杆菌的同源蛋白。总体而言，这些数据表明，副猪隐孢子虫和巴氏杆菌科其他成员一样，具有获取铁的TonB区的遗传元件，并且该区域下编码的转铁蛋白结合蛋白在铁可利用性受限的情况下上调。(C)2005年欧洲微生物学会联合会。爱思唯尔出版，版权所有。
Haemophilus parasuis is the causative agent of Glasser's disease, which is responsible for considerable economic losses in the pig-rearing industry. The aim of the study reported here was the identification, sequencing and molecular characterization of the TonB region that includes tonB. exbBD, and tbpBA genes in H. parasuis. In addition, two fusion proteins were generated. One of them (pGEX-6P-1-GST-TbpB) contained the first 501 amino acids of H. parasuis TbpB protein, while the second (pBAD-Thio-TbpB-V5-His) included the first 102 amino acids of H. parasuis TbpB N-terminus domain. A panel of 14 hybridomas secreting monoclonal antibodies was raised against the two recombinant TbpB fusion proteins. Furthermore, to assess whether the expression of the H. parasuis ExbB, TbpB, and TbpA proteins was upregulated under conditions of restricted availability of iron, a rabbit polyclonal antibody against H. parasuis TbpB-His fusion protein was produced. A rabbit polyclonal antibody against serotype 7 of Actinobacillus pleuropneumoniae ExbB and TbpA proteins was also used for the detection of the homologous proteins in H. parasuis. Overall, the data indicate that H. parasuis, like other members of the Pasteurellaceae family, possesses the genetic elements of the TonB region for iron acquisition and the transferrin-binding proteins encoded under this region are upregulated under restricted iron availability. (c) 2005 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.