Sato S, Ozeki Y, Kawaguchi A
Department of Biology, College of Arts and Sciences, University of Tokyo.
J Biochem. 1994 Feb;115(2):286-92. doi: 10.1093/oxfordjournals.jbchem.a124330.
Transcription of the two clustered genes, faoA and faoB, for the multienzyme complex for fatty acid beta-oxidation (HDT) from the Gram-negative bacterium, Pseudomonas fragi, was investigated. Northern blot hybridization and primer extension analysis indicated that these genes comprised an operon, giving a 3.6 kb mRNA. Transcription of the 3.6 kb faoAB mRNA was induced by palmitic acid. Using the lacZ gene as a reporter gene for a promoter activity search, the essential region for induction of the faoAB gene transcription by palmitic acid was mapped at the 331 bp region just upstream of the transcription initiation site. Deletion of the 136 bp SalI-NheI fragment just downstream of the transcription initiation site caused the constitutive expression of the faoA-lacZ fusion protein in P. fragi, suggesting that there is a regulatory element interacting with the transcription repressor.
对革兰氏阴性菌脆弱拟杆菌中负责脂肪酸β-氧化多酶复合体(HDT)的两个成簇基因faoA和faoB的转录进行了研究。Northern印迹杂交和引物延伸分析表明,这些基因组成一个操纵子,产生一条3.6 kb的mRNA。3.6 kb的faoAB mRNA转录由棕榈酸诱导。使用lacZ基因作为报告基因进行启动子活性搜索,棕榈酸诱导faoAB基因转录的必需区域定位在转录起始位点上游331 bp处。转录起始位点下游紧挨着的136 bp SalI-NheI片段缺失导致faoA-lacZ融合蛋白在脆弱拟杆菌中组成型表达,这表明存在一个与转录阻遏物相互作用的调控元件。