IDENTIFICATION OF GENES AND GENE-PRODUCTS NECESSARY FOR BACTERIAL BIOLUMINESCENCE
IDENTIFICATION OF GENES AND GENE-PRODUCTS NECESSARY FOR BACTERIAL BIOLUMINESCENCE
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DOI：
10.1073/pnas.81.13.4154
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发表时间：
1984-01-01
期刊：
影响因子：
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通讯作者：
SILVERMAN, M
中科院分区：
文献类型：
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作者：
ENGEBRECHT, J;SILVERMAN, M
最近通过克隆海洋细菌费氏弧菌的基因，在大肠杆菌中实现了发光的表达。杂交质粒上的一个DNA片段编码光产生所必需的调节功能和酶活性。报告了对位于该重组质粒上的发光基因(Lux)进行遗传分析的结果。Lux基因突变是通过羟胺处理产生的，这些突变在线性图谱上通过反式互补和一系列极性转座子插入到其他质粒上而被排序。通过在反式表达载体上互补lux基因缺陷，确定了lux基因。杂交质粒也被用来指导大肠杆菌微细胞系统中多肽的合成。通过互补分析和微细胞编程实验，确定了7个Lux基因及其相应的基因产物。这些基因按照它们在线性地图上的位置顺序，基因产物的表观分子量是LuxR(27,000)、Luxi(25,000)、LuxC(53,000)、LuxD(33,000)、LuxA(40,000)、LuxB(38,000)和LuxE(42,000)。根据含有突变质粒的大肠杆菌的发光表型，功能被分配给这些基因：LuxA、LuxB、LuxC、LuxD和LuxE编码发光的酶，LuxR和Luxi编码调节功能。
Expression of luminescence in Escherichia coli was recently achieved by cloning genes from the marine bacterium Vibrio fischeri. One DNA fragment on a hybrid plasmid encoded regulatory functions and enzymatic activities necessary for light production. The results of a genetic analysis to identify the luminescence genes (lux) that reside on this recombinant plasmid are reported. The lux gene mutations were generated by hydroxylamine treatment, and these mutations were ordered on a linear map by complementation in trans with a series of polar transposon insertions on other plasmids. The lux genes were defined by complementation of lux gene defects on pairs of plasmids in trans in E. coli. Hybrid plasmids were also used to direct the synthesis of polypeptides in the E. coli minicell system. Seven lux genes and the corresponding gene products were identified from the complementation analysis and the minicell programming experiments. These genes, in the order of their position on a linear map, and the apparent MW of the gene products are luxR (27,000), luxI (25,000), luxC (53,000), luxD (33,000), luxA (40,000), luxB (38,000) and luxE (42,000). From the luminescence phenotypes of E. coli containing mutant plasmids, functions were assigned to these genes: luxA, luxB, luxC, luxD and luxE encode enzymes for light production, and luxR and luxI encode regulatory functions.