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杜克雷嗜血杆菌孔蛋白OmpP2A和OmpP2B的差异表达

Differential expression of porins OmpP2A and OmpP2B of Haemophilus ducreyi.

作者信息

Prather Derrick T, Bains Manjeet, Hancock Robert E W, Filiatrault Melanie J, Campagnari Anthony A

机构信息

Department of Microbiology and Immunology, State University of New York at Buffalo, 14214, USA.

出版信息

Infect Immun. 2004 Nov;72(11):6271-8. doi: 10.1128/IAI.72.11.6271-6278.2004.

Abstract

Haemophilus ducreyi is a strict human pathogen and the causative agent of the sexually transmitted disease chancroid. The genome of the human-passaged strain of H. ducreyi (35000HP) contains two homologous genes whose protein products have estimated molecular masses of 46 and 43 kDa. A comparative analysis of the deduced amino acid sequences revealed that these proteins share 27 to 33% identity to the outer membrane protein P2 (OmpP2), a major porin of Haemophilus influenzae. Therefore, these proteins have been designated OmpP2A and OmpP2B, respectively. The detection of ompP2A and ompP2B transcripts by reverse transcriptase PCR indicated that these genes were independently transcribed in H. ducreyi 35000HP. Western blot analysis of outer membrane proteins isolated from a geographically diverse collection of H. ducreyi clinical isolates revealed that OmpP2A and OmpP2B were differentially expressed among these strains. Although PCR analysis suggested that ompP2A and ompP2B were conserved among the strains tested, the differential expression observed was due to nucleotide additions and partial gene deletions. Purified OmpP2A and OmpP2B were isolated under nondenaturing conditions, and subsequent analysis demonstrated that these two proteins exhibited porin activity. OmpP2A and OmpP2B are the first porins described for H. ducreyi.

摘要

杜克雷嗜血杆菌是一种严格的人类病原体,也是性传播疾病软下疳的病原体。杜克雷嗜血杆菌人传代菌株(35000HP)的基因组包含两个同源基因,其蛋白质产物的估计分子量分别为46 kDa和43 kDa。对推导的氨基酸序列进行比较分析发现,这些蛋白质与流感嗜血杆菌的主要孔蛋白外膜蛋白P2(OmpP2)有27%至33%的同一性。因此,这些蛋白质分别被命名为OmpP2A和OmpP2B。通过逆转录酶PCR检测ompP2A和ompP2B转录本表明,这些基因在杜克雷嗜血杆菌35000HP中是独立转录的。对从不同地理区域收集的杜克雷嗜血杆菌临床分离株中分离的外膜蛋白进行的蛋白质印迹分析表明,OmpP2A和OmpP2B在这些菌株中差异表达。尽管PCR分析表明ompP2A和ompP2B在所测试的菌株中是保守的,但观察到的差异表达是由于核苷酸添加和部分基因缺失所致。在非变性条件下分离纯化了OmpP2A和OmpP2B,随后的分析表明这两种蛋白质具有孔蛋白活性。OmpP2A和OmpP2B是首次描述的杜克雷嗜血杆菌孔蛋白。

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