Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 Jul;81(13):4154–4158. doi: 10.1073/pnas.81.13.4154

Identification of genes and gene products necessary for bacterial bioluminescence.

J Engebrecht, M Silverman
PMCID: PMC345387  PMID: 6377310

Abstract

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. We report the results of a genetic analysis to identify the luminescence genes (lux) that reside on this recombinant plasmid. 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. 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 programing experiments. These genes, in the order of their position on a linear map, and the apparent molecular weights 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.

Full text

PDF
4154

Images in this article

4156Image
on p.4156
4157Image
on p.4157

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baumann P., Baumann L. Biology of the marine enterobacteria: genera Beneckea and Photobacterium. Annu Rev Microbiol. 1977;31:39–61. doi: 10.1146/annurev.mi.31.100177.000351. [DOI] [PubMed] [Google Scholar]
  2. Belas R., Mileham A., Cohn D., Hilman M., Simon M., Silverman M. Bacterial bioluminescence: isolation and expression of the luciferase genes from Vibrio harveyi. Science. 1982 Nov 19;218(4574):791–793. doi: 10.1126/science.10636771. [DOI] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  5. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohn D. H., Ogden R. C., Abelson J. N., Baldwin T. O., Nealson K. H., Simon M. I., Mileham A. J. Cloning of the Vibrio harveyi luciferase genes: use of a synthetic oligonucleotide probe. Proc Natl Acad Sci U S A. 1983 Jan;80(1):120–123. doi: 10.1073/pnas.80.1.120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eberhard A., Burlingame A. L., Eberhard C., Kenyon G. L., Nealson K. H., Oppenheimer N. J. Structural identification of autoinducer of Photobacterium fischeri luciferase. Biochemistry. 1981 Apr 28;20(9):2444–2449. doi: 10.1021/bi00512a013. [DOI] [PubMed] [Google Scholar]
  8. Engebrecht J., Nealson K., Silverman M. Bacterial bioluminescence: isolation and genetic analysis of functions from Vibrio fischeri. Cell. 1983 Mar;32(3):773–781. doi: 10.1016/0092-8674(83)90063-6. [DOI] [PubMed] [Google Scholar]
  9. Gunsalus-Miguel A., Meighen E. A., Nicoli M. Z., Nealson K. H., Hastings J. W. Purification and properties of bacterial luciferases. J Biol Chem. 1972 Jan 25;247(2):398–404. [PubMed] [Google Scholar]
  10. Mandel M., Higa A. Calcium-dependent bacteriophage DNA infection. J Mol Biol. 1970 Oct 14;53(1):159–162. doi: 10.1016/0022-2836(70)90051-3. [DOI] [PubMed] [Google Scholar]
  11. Matsumura P., Silverman M., Simon M. Synthesis of mot and che gene products of Escherichia coli programmed by hybrid ColE1 plasmids in minicells. J Bacteriol. 1977 Dec;132(3):996–1002. doi: 10.1128/jb.132.3.996-1002.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Nealson K. H. Autoinduction of bacterial luciferase. Occurrence, mechanism and significance. Arch Microbiol. 1977 Feb 4;112(1):73–79. doi: 10.1007/BF00446657. [DOI] [PubMed] [Google Scholar]
  13. Nealson K. H., Hastings J. W. Bacterial bioluminescence: its control and ecological significance. Microbiol Rev. 1979 Dec;43(4):496–518. doi: 10.1128/mr.43.4.496-518.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rodriguez A., Riendeau D., Meighen E. Purification of the acyl coenzyme A reductase component from a complex responsible for the reduction of fatty acids in bioluminescent bacteria. Properties and acyltransferase activity. J Biol Chem. 1983 Apr 25;258(8):5233–5237. [PubMed] [Google Scholar]
  15. Silerman M., Matsumura P., Draper R., Edwards S., Simon M. I. Expression of flagellar genes carried by bacteriophage lambda. Nature. 1976 May 20;261(5557):248–250. doi: 10.1038/261248a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES