Martin M, Showalter R, Silverman M
Agouron Institute, La Jolla, California 92037.
J Bacteriol. 1989 May;171(5):2406-14. doi: 10.1128/jb.171.5.2406-2414.1989.
Mutagenesis with transposon mini-Mulac was used to identify loci containing genes for bioluminescence (lux) in the marine bacterium Vibrio harveyi. Transposon insertions which resulted in a Lux- phenotype were mapped to two unlinked regions of the genome. Region I contained the luxCDABE operon which was previously shown to encode the enzymes luciferase and fatty acid reductase, which are required for light production. The other locus, region II, which was identified for the first time in this study, appeared to have a regulatory function. In Northern blot analysis of mRNA from mutants with defects in this region, no transcription from the luxCDABE operon could be detected. Strains with transposon-generated lux::lacZ gene fusions were used to analyze control of the transcription of these regions. Expression of luminescence in the wild type was strongly influenced by the density of the culture, and in strains with the lacZ indicator gene coupled to the luxCDABE operon, beta-galactosidase synthesis was density dependent. So, transcription of this operon is responsive to a density-sensing mechanism. However, beta-galactosidase synthesis in strains with lacZ fused to the region II transcriptional unit did not respond to cell density. The organization and regulation of the lux genes of V. harveyi are discussed, particularly with regard to the contrasts observed with the lux system of the fish light-organ symbiont Vibrio fischeri.
利用转座子mini-Mulac进行诱变,以鉴定海洋细菌哈维氏弧菌中含有生物发光(lux)基因的位点。导致Lux-表型的转座子插入被定位到基因组的两个不连锁区域。区域I包含luxCDABE操纵子,该操纵子先前已被证明编码荧光素酶和脂肪酸还原酶,这些酶是发光所必需的。另一个位点,区域II,是在本研究中首次鉴定出来的,似乎具有调控功能。在对该区域有缺陷的突变体的mRNA进行Northern印迹分析时,未检测到来自luxCDABE操纵子的转录。利用转座子产生的lux::lacZ基因融合菌株来分析这些区域转录的调控。野生型中发光的表达受到培养物密度的强烈影响,并且在lacZ指示基因与luxCDABE操纵子偶联的菌株中,β-半乳糖苷酶的合成是密度依赖性的。因此,该操纵子的转录对密度感应机制有反应。然而,lacZ与区域II转录单元融合的菌株中β-半乳糖苷酶的合成对细胞密度没有反应。本文讨论了哈维氏弧菌lux基因的组织和调控,特别是与鱼类发光器官共生菌费氏弧菌的lux系统所观察到的差异有关。