Günel-Ozcan A, Brown K A, Allen A G, Maskell D J
Department of Biochemistry, Imperial College of Science, Technology & Medicine, London, SW7 2AY, U.K.
Microb Pathog. 1997 Nov;23(5):311-6. doi: 10.1006/mpat.1997.0157.
The aroB gene of Salmonella typhimurium, encoding dehydroquinate synthase, has been cloned into pUC19 and the DNA sequence determined. The aroB gene was isolated from a cosmid gene bank by complementation of an Escherichia coli aroB mutant and screening by Southern blot analysis. The nucleotide sequence of the S. typhimurium aroB gene revealed the presence of an open reading frame, encoding a protein of 362 amino acids with a calculated molecular mass of 38696 Daltons. The amino acid sequence of S. typhimurium dehydroquinate synthase is nearly identical to the E. coli homologue and shows high homology with other aroB gene products from other organisms. Subsequently, a stable insertional mutation in aroB was introduced into the wild-type S. typhimurium C5 strain. This mutant was auxotrophic for aromatic compounds. Infection of BALB/c mice with this mutant demonstrated attenuation comparable to other S. typhimurium mutants unable to biosynthesize aromatic amino acids.
鼠伤寒沙门氏菌的aroB基因编码脱氢奎尼酸合酶,已被克隆到pUC19中并测定了DNA序列。通过对大肠杆菌aroB突变体进行互补并利用Southern印迹分析进行筛选,从黏粒基因文库中分离出aroB基因。鼠伤寒沙门氏菌aroB基因的核苷酸序列显示存在一个开放阅读框,编码一种由362个氨基酸组成的蛋白质,计算分子量为38696道尔顿。鼠伤寒沙门氏菌脱氢奎尼酸合酶的氨基酸序列与大肠杆菌的同源物几乎相同,并且与来自其他生物体的其他aroB基因产物具有高度同源性。随后,在野生型鼠伤寒沙门氏菌C5菌株中引入了aroB的稳定插入突变。该突变体对芳香族化合物营养缺陷。用该突变体感染BALB/c小鼠显示出与其他无法生物合成芳香族氨基酸的鼠伤寒沙门氏菌突变体相当的减毒效果。