Choe M, Reznikoff W S
Department of Biochemistry, University of Wisconsin, Madison 53706.
J Bacteriol. 1991 Oct;173(19):6139-46. doi: 10.1128/jb.173.19.6139-6146.1991.
Genes that are expressed under anaerobic conditions were identified by operon fusion techniques with a hybrid bacteriophage of lambda and Mu, lambda placMu53, which creates transcriptional fusions to lacZY. Cells were screened for anaerobic expression on XG medium. Nine strains were selected, and the insertion point of the hybrid phage in each strain was mapped on the Escherichia coli chromosome linkage map. The anaerobic and aerobic expression levels of these genes were measured by beta-galactosidase assays in different medium conditions and in the presence of three regulatory mutations (fnr, narL, and rpoN). The anaerobically expressed genes (aeg) located at minute 99 (aeg-99) and 75 (aeg-75) appeared to be partially regulated by fnr, and aeg-93 is tightly regulated by fnr. aeg-60 requires a functional rpoN gene for its anaerobic expression. aeg-46.5 is repressed by narL. aeg-65A and aeg-65C are partially controlled by fnr but only in media containing nitrate or fumarate. aeg-47.5 and aeg-48.5 were found to be anaerobically induced only in rich media. The effects of a narL mutation on aeg-46.5 expression were observed in all medium conditions regardless of the presence or absence of nitrate. This suggests that narL has a regulatory function in the absence of exogenously added nitrate.
利用λ噬菌体与Mu噬菌体的杂交噬菌体λplacMu53通过操纵子融合技术鉴定了在厌氧条件下表达的基因,该杂交噬菌体可产生与lacZY的转录融合。在XG培养基上筛选细胞的厌氧表达。选择了9个菌株,并将每个菌株中杂交噬菌体的插入点定位在大肠杆菌染色体连锁图谱上。在不同培养基条件下以及存在三种调节突变(fnr、narL和rpoN)的情况下,通过β-半乳糖苷酶测定法测量这些基因的厌氧和好氧表达水平。位于99分钟处(aeg-99)和75分钟处(aeg-75)的厌氧表达基因似乎部分受fnr调节,而aeg-93受fnr严格调节。aeg-60的厌氧表达需要功能性的rpoN基因。aeg-46.5受narL抑制。aeg-65A和aeg-65C仅在含有硝酸盐或富马酸盐的培养基中部分受fnr控制。发现aeg-47.5和aeg-48.5仅在丰富培养基中被厌氧诱导。无论是否存在硝酸盐,在所有培养基条件下均观察到narL突变对aeg-46.5表达的影响。这表明narL在未添加外源硝酸盐的情况下具有调节功能。