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奇异变形杆菌环境温度菌毛:aft基因簇的克隆与核苷酸序列分析

Proteus mirabilis ambient-temperature fimbriae: cloning and nucleotide sequence of the aft gene cluster.

作者信息

Massad G, Fulkerson J F, Watson D C, Mobley H L

机构信息

Division of Infectious Diseases, University of Maryland School of Medicine, Baltimore 21201, USA.

出版信息

Infect Immun. 1996 Oct;64(10):4390-5. doi: 10.1128/iai.64.10.4390-4395.1996.

Abstract

Uropathogenic Proteus mirabilis produces at least four types of fimbriae. Amino acid sequences from two peptides, derived by tryptic digestion of the structural subunit of one type of these fimbriae, the ambient-temperature fimbriae, were determined: NVVPGQPSSTQ and LIEGENQLNYNA. PCR primers, based on these sequences and that of the N terminus, were used to amplify a 359-bp fragment. A cosmid clone, isolated from a P. mirabilis genomic library by hybridization with the 359-bp PCR product, was used to determine the nucleotide sequence of the atf gene cluster. A 3,903-bp region encodes three polypeptides: AtfA, the structural subunit; AtfB, the chaperone; and AtfC, the outer membrane molecular usher. No fimbria-related genes are evident either 5' or 3' to the three contiguous genes. AtfA demonstrates significant amino acid sequence identity with type 1 major fimbrial subunits of several enteric species. The 359-bp PCR product hybridized strongly with all Proteus isolates (n = 9) and 25% of 355 Escherichia coli isolates but failed to hybridize with any of 26 isolates among nine other uropathogenic species. Ambient-temperature fimbriae of P. mirabilis may represent a novel type of fimbriae of enteric species.

摘要

尿路致病性奇异变形杆菌可产生至少四种类型的菌毛。对其中一种菌毛(即常温菌毛)的结构亚基进行胰蛋白酶消化后得到的两个肽段的氨基酸序列已被确定:NVVPGQPSSTQ和LIEGENQLNYNA。基于这些序列以及N端序列设计的PCR引物,被用于扩增一段359 bp的片段。通过与359 bp的PCR产物杂交,从奇异变形杆菌基因组文库中分离出一个黏粒克隆,用于确定atf基因簇的核苷酸序列。一个3903 bp的区域编码三种多肽:结构亚基AtfA、伴侣蛋白AtfB和外膜分子伴侣AtfC。在这三个相邻基因的5'端或3'端均未发现明显的菌毛相关基因。AtfA与几种肠道菌的1型主要菌毛亚基具有显著的氨基酸序列同一性。359 bp的PCR产物与所有奇异变形杆菌分离株(n = 9)以及355株大肠杆菌分离株中的25%发生强烈杂交,但与其他九种尿路致病性菌种的26株分离株均未发生杂交。奇异变形杆菌的常温菌毛可能代表了一种新型的肠道菌菌毛。

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