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来自半乳糖阳性细菌唾液链球菌以及系统发育相关的半乳糖阴性细菌嗜热链球菌的半乳糖和乳糖基因:gal基因产物的组织、序列、转录及活性

Galactose and lactose genes from the galactose-positive bacterium Streptococcus salivarius and the phylogenetically related galactose-negative bacterium Streptococcus thermophilus: organization, sequence, transcription, and activity of the gal gene products.

作者信息

Vaillancourt Katy, Moineau Sylvain, Frenette Michel, Lessard Christian, Vadeboncoeur Christian

机构信息

Groupe de Recherche en Ecologie Buccale, Département de Biochimie et de Microbiologie, Faculté des Sciences et de Génie and Faculté de Médecine Dentaire, Université Laval, Québec, Québec, Canada G1K 7P4.

出版信息

J Bacteriol. 2002 Feb;184(3):785-93. doi: 10.1128/JB.184.3.785-793.2002.

Abstract

Streptococcus salivarius is a lactose- and galactose-positive bacterium that is phylogenetically closely related to Streptococcus thermophilus, a bacterium that metabolizes lactose but not galactose. In this paper, we report a comparative characterization of the S. salivarius and S. thermophilus gal-lac gene clusters. The clusters have the same organization with the order galR (codes for a transcriptional regulator and is transcribed in the opposite direction), galK (galactokinase), galT (galactose-1-P uridylyltransferase), galE (UDP-glucose 4-epimerase), galM (galactose mutarotase), lacS (lactose transporter), and lacZ (beta-galactosidase). An analysis of the nucleotide sequence as well as Northern blotting and primer extension experiments revealed the presence of four promoters located upstream from galR, the gal operon, galM, and the lac operon of S. salivarius. Putative promoters with virtually identical nucleotide sequences were found at the same positions in the S. thermophilus gal-lac gene cluster. An additional putative internal promoter at the 3' end of galT was found in S. thermophilus but not in S. salivarius. The results clearly indicated that the gal-lac gene cluster was efficiently transcribed in both species. The Shine-Dalgarno sequences of galT and galE were identical in both species, whereas the ribosome binding site of S. thermophilus galK differed from that of S. salivarius by two nucleotides, suggesting that the S. thermophilus galK gene might be poorly translated. This was confirmed by measurements of enzyme activities.

摘要

唾液链球菌是一种乳糖和半乳糖阳性细菌,在系统发育上与嗜热链球菌密切相关,嗜热链球菌是一种能代谢乳糖但不能代谢半乳糖的细菌。在本文中,我们报告了唾液链球菌和嗜热链球菌gal-lac基因簇的比较特征。这些基因簇具有相同的组织形式,顺序为galR(编码转录调节因子,转录方向相反)、galK(半乳糖激酶)、galT(半乳糖-1-磷酸尿苷酰转移酶)、galE(UDP-葡萄糖4-差向异构酶)、galM(半乳糖变旋酶)、lacS(乳糖转运蛋白)和lacZ(β-半乳糖苷酶)。对核苷酸序列的分析以及Northern印迹和引物延伸实验表明,在唾液链球菌的galR、gal操纵子、galM和lac操纵子上游存在四个启动子。在嗜热链球菌gal-lac基因簇的相同位置发现了核苷酸序列几乎相同的推定启动子。在嗜热链球菌galT的3'端发现了一个额外的推定内部启动子,而在唾液链球菌中未发现。结果清楚地表明,gal-lac基因簇在这两个物种中均能有效转录。galT和galE的Shine-Dalgarno序列在两个物种中相同,而嗜热链球菌galK的核糖体结合位点与唾液链球菌的核糖体结合位点有两个核苷酸的差异,这表明嗜热链球菌galK基因的翻译可能较差。酶活性测量证实了这一点。

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