Mancini J A, Boylan M, Soly R R, Graham A F, Meighen E A
Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
J Biol Chem. 1988 Oct 5;263(28):14308-14.
The organization of the lux structural genes (A-E) in Photobacterium phosphoreum has been determined and a new gene designated as luxF discovered. The P. phosphoreum luminescence system was cloned into Escherichia coli using a pBR322 vector and identified by cross-hybridization with Vibrio fischeri lux DNA. The lux genes were located by specific expression of P. phosphoreum DNA fragments in the T7-phage polymerase/promoter system in E. coli and identification of the labeled polypeptide products. The luxA and luxB gene products (luciferase subunits) were shown to catalyze light emission in the presence of FMNH2, O2, and aldehyde. The luxC, luxD, and luxE gene products (fatty acid reductase subunits) responsible for aldehyde biosynthesis could be specifically acylated with 3H-labeled fatty acids. The order of the lux genes in P. phosphoreum was found to be luxCDABFE with luxF coding for a new polypeptide of 26 kDa. The presence of a new gene in the P. phosphoreum luminescence system between luxB and luxE as compared to the organization of the lux structural gene in V. fischeri and Vibrio harveyi (luxCDABE) demonstrates that the luminescent systems in the marine bacteria have significantly diverged. The discovery of the luxF gene provides the basis for elucidating the role of its gene product in the expression of luminescence in different marine bacteria.
已经确定了发光杆菌中lux结构基因（A - E）的组织方式，并发现了一个新的基因，命名为luxF。利用pBR322载体将发光杆菌的发光系统克隆到大肠杆菌中，并通过与费氏弧菌lux DNA的交叉杂交进行鉴定。通过在大肠杆菌的T7噬菌体聚合酶/启动子系统中特异性表达发光杆菌DNA片段并鉴定标记的多肽产物来定位lux基因。已证明luxA和luxB基因产物（荧光素酶亚基）在存在FMNH2、O2和醛的情况下催化发光。负责醛生物合成的luxC、luxD和luxE基因产物（脂肪酸还原酶亚基）可以用3H标记的脂肪酸进行特异性酰化。发现发光杆菌中lux基因的顺序为luxCDABFE，其中luxF编码一种26 kDa的新多肽。与费氏弧菌和哈维氏弧菌（luxCDABE）中lux结构基因的组织方式相比，发光杆菌发光系统中在luxB和luxE之间存在一个新基因，这表明海洋细菌中的发光系统已经有了显著的分化。luxF基因的发现为阐明其基因产物在不同海洋细菌发光表达中的作用提供了基础。
