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乌普萨拉弯曲杆菌铁摄取调节蛋白(fur)的特性及其在弯曲杆菌基因组高度保守区域的定位

Characterization of Campylobacter upsaliensis fur and its localization in a highly conserved region of the Campylobacter genome.

作者信息

Bourke B, al Rashid S T, Bingham H L, Chan V L

机构信息

Division of Gastroenterology, Hospital for Sick Children, University of Toronto, Ontario, Canada.

出版信息

Gene. 1996 Dec 12;183(1-2):219-24. doi: 10.1016/s0378-1119(96)00562-8.

Abstract

Despite increasing recognition of the importance of Campylobacter upsaliensis in human disease little is known about either the virulence properties or genetics of this enteric pathogen. The complete coding sequence of a C. upsaliensis gene has yet to be published. We have cloned and sequenced the complete iron-uptake regulatory (fur) gene from the type strain of this species. The C. upsaliensis fur homolog was isolated from a genomic library of C. upsaliensis ATCC 43954 constructed in phage lambdaGEM-11. The open reading frame identified encodes a polypeptide consisting of 156 amino acids. The 5'-flanking region of the C. upsaliensis fur gene contains 3 putative Fur-binding sequences and two catabolite activator-binding sequences indicating the potential for autogenous and cAMP-mediated regulation, respectively. Primer extension analysis identified a single transcription start site 262 nt upstream from the AUG initiation codon. Sequence analysis indicates that the Fur protein of C. upsaliensis is highly homologous (87% amino acid identity) to Campylobacter jejuni Fur. Furthermore, the arrangement of the lysS and glyA genes downstream of fur is precisely conserved in both C. upsaliensis ATCC 43954 and C. jejuni TGH9011. Using the polymerase chain reaction close linkage of fur with lysS and glyA was also observed among multiple isolates of C. upsaliensis, C. jejuni and C. coli suggesting a possible functional relevance for this conserved genetic arrangement in campylobacteria.

摘要

尽管人们越来越认识到乌普萨拉弯曲菌在人类疾病中的重要性,但对于这种肠道病原体的毒力特性或遗传学却知之甚少。乌普萨拉弯曲菌基因的完整编码序列尚未公布。我们已经从该物种的模式菌株中克隆并测序了完整的铁摄取调节(fur)基因。乌普萨拉弯曲菌fur同源物是从构建于噬菌体lambdaGEM - 11中的乌普萨拉弯曲菌ATCC 43954基因组文库中分离出来的。鉴定出的开放阅读框编码一个由156个氨基酸组成的多肽。乌普萨拉弯曲菌fur基因的5'侧翼区域包含3个推定的Fur结合序列和两个分解代谢物激活剂结合序列,分别表明了自体调节和cAMP介导调节的可能性。引物延伸分析确定了一个位于AUG起始密码子上游262 nt处的单一转录起始位点。序列分析表明,乌普萨拉弯曲菌的Fur蛋白与空肠弯曲菌Fur高度同源(氨基酸同一性为87%)。此外,fur下游的lysS和glyA基因的排列在乌普萨拉弯曲菌ATCC 43954和空肠弯曲菌TGH9011中都精确保守。使用聚合酶链反应,在乌普萨拉弯曲菌、空肠弯曲菌和大肠弯曲菌的多个分离株中也观察到fur与lysS和glyA的紧密连锁,这表明这种保守的基因排列在弯曲菌中可能具有功能相关性。

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