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嗜酸乳杆菌L-(+)-乳酸脱氢酶基因的克隆、核苷酸序列及转录分析

Cloning, nucleotide sequence, and transcriptional analysis of the Pediococcus acidilactici L-(+)-lactate dehydrogenase gene.

作者信息

Garmyn D, Ferain T, Bernard N, Hols P, Delcour J

机构信息

Laboratoire de Génétique Moléculaire, Université Catholique de Louvain, Belgium.

出版信息

Appl Environ Microbiol. 1995 Jan;61(1):266-72. doi: 10.1128/aem.61.1.266-272.1995.

Abstract

Recombinant plasmids containing the Pediococcus acidilactici L-(+)-lactate dehydrogenase gene (ldhL) were isolated by complementing for growth under anaerobiosis of an Escherichia coli lactate dehydrogenase-pyruvate formate lyase double mutant. The nucleotide sequence of the ldhL gene predicted a protein of 323 amino acids showing significant similarity with other bacterial L-(+)-lactate dehydrogenases and especially with that of Lactobacillus plantarum. The ldhL transcription start points in P. acidilactici were defined by primer extension, and the promoter sequence was identified as TCAAT-(17 bp)-TATAAT. This sequence is closely related to the consensus sequence of vegetative promoters from gram-positive bacteria as well as from E. coli. Northern analysis of P. acidilactici RNA showed a 1.1-kb ldhL transcript whose abundance is growth rate regulated. These data, together with the presence of a putative rho-independent transcriptional terminator, suggest that ldhL is expressed as a monocistronic transcript in P. acidilactici.

摘要

通过在大肠杆菌乳酸脱氢酶-丙酮酸甲酸裂解酶双突变体的厌氧生长条件下进行互补,分离出了含有嗜酸乳杆菌L-(+)-乳酸脱氢酶基因(ldhL)的重组质粒。ldhL基因的核苷酸序列预测其编码一个由323个氨基酸组成的蛋白质,该蛋白质与其他细菌的L-(+)-乳酸脱氢酶具有显著相似性,尤其与植物乳杆菌的L-(+)-乳酸脱氢酶相似。通过引物延伸确定了嗜酸乳杆菌中ldhL的转录起始点,启动子序列被鉴定为TCAAT-(17bp)-TATAAT。该序列与革兰氏阳性菌以及大肠杆菌的营养型启动子的共有序列密切相关。对嗜酸乳杆菌RNA的Northern分析显示有一个1.1kb的ldhL转录本,其丰度受生长速率调控。这些数据,连同推定的不依赖ρ因子的转录终止子的存在,表明ldhL在嗜酸乳杆菌中以单顺反子转录本的形式表达。

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