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嗜肺军团菌铁摄取调节蛋白基因的克隆与测序

Cloning and sequencing of the Legionella pneumophila fur gene.

作者信息

Hickey E K, Cianciotto N P

机构信息

Department of Microbiology-Immunology, Northwestern University, Chicago, IL 60611.

出版信息

Gene. 1994 May 27;143(1):117-21. doi: 10.1016/0378-1119(94)90615-7.

Abstract

Iron is required for the intracellular and extracellular growth of Legionella pneumophila (Lp). In addition, variations in iron levels may serve as a signal for changes in gene expression. In a number of bacterial pathogens, the regulation of gene expression by iron is usually mediated by the Fur (ferric uptake regulation) repressor protein. Through complementation of an Escherichia coli fur mutation and nucleotide sequence analysis, we have cloned and characterized the Lp fur gene. Lp fur encoded a 15.0-kDa protein whose repressive activity was, as expected, highest in bacteria grown in iron-rich media. Computer analysis determined that Lp Fur had an amino-acid identity of over 54% and a similarity of over 72% to the Fur of E. coli, Yersinia pestis, Vibrio species and Pseudomonas aeruginosa. The promoter region of Lp fur contained sequences homologous to the Fur-binding site, suggesting that fur is autoregulated in Lp. Finally, Southern blot hybridizations demonstrated that fur is conserved among Lp strains and Legionella species.

摘要

嗜肺军团菌(Lp)在细胞内和细胞外生长均需要铁。此外,铁水平的变化可能作为基因表达改变的信号。在许多细菌病原体中,铁对基因表达的调控通常由Fur(铁摄取调节)阻遏蛋白介导。通过对大肠杆菌fur突变体的互补和核苷酸序列分析,我们克隆并鉴定了Lp fur基因。Lp fur编码一种15.0 kDa的蛋白质,其阻遏活性在富含铁的培养基中生长的细菌中如预期的那样最高。计算机分析确定,Lp Fur与大肠杆菌、鼠疫耶尔森菌、弧菌属和铜绿假单胞菌的Fur具有超过54%的氨基酸同一性和超过72%的相似性。Lp fur的启动子区域包含与Fur结合位点同源的序列,表明fur在Lp中是自我调节的。最后,Southern印迹杂交表明fur在Lp菌株和军团菌属中是保守的。

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