Eraso J M, Weinstock G M
Department of Biochemistry and Molecular Biology, University of Texas Medical School, Houston 77225.
J Bacteriol. 1992 Aug;174(15):5101-9. doi: 10.1128/jb.174.15.5101-5109.1992.
Expression of the cea gene, which is carried by the ColE1 plasmid and which encodes colicin E1, was found to be greatly increased when the cells were grown anaerobically. By using cea-lacZ fusions to quantitate expression, aerobic levels were found to be only a few percent of the anaerobic levels. The anaerobic increase in expression was observed both in protein and in operon fusions, indicating that its regulation occurred at the level of transcription. It was also found to require a functional fnr gene and to occur when the cea-lacZ fusion was present as a single copy in the bacterial chromosome instead of in the multicopy ColE1 plasmid. Anaerobic expression was regulated by the SOS response and catabolite repression as is aerobic expression. The start site of the mRNA produced under anaerobic conditions was mapped by primer extension and found to be the same as the start for mRNA produced under aerobic conditions. These observations show that the cea gene is anaerobically regulated and that the Fnr protein is a positive regulator of transcription of this gene.
由ColE1质粒携带并编码大肠杆菌素E1的cea基因,在细胞厌氧生长时其表达量显著增加。通过使用cea - lacZ融合体来定量表达,发现需氧水平仅为厌氧水平的百分之几。在蛋白质融合体和操纵子融合体中均观察到厌氧条件下表达量的增加,这表明其调控发生在转录水平。还发现这需要一个功能性的fnr基因,并且当cea - lacZ融合体以单拷贝形式存在于细菌染色体而非多拷贝的ColE1质粒中时才会发生。厌氧表达与需氧表达一样,受SOS反应和分解代谢物阻遏的调控。通过引物延伸法确定了厌氧条件下产生的mRNA的起始位点,发现其与需氧条件下产生的mRNA的起始位点相同。这些观察结果表明cea基因受到厌氧调控,并且Fnr蛋白是该基因转录的正调控因子。