Skip to main content
Log in

The use of the luxA gene of the bacterial luciferase operon as a reporter gene

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Bacterial luciferase can be assayed rapidly and with high sensitivity both in vivo and in vitro. Here we demonstrate that the N-terminal hydrophobic domain of the α catalytic subunit of the luciferase enzyme is indispensable for enzyme activity, although N-terminal translational fusions with full luciferase activity can be obtained. Bacterial luciferase is therefore ideally suited as a reporter enzyme for gene fusion experiments. A list of vectors for the convenient use of the luciferase marker genes to monitor gene expression in vivo are presented.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Baldwin TO, Berends T, Bunch TA, Holzman TF, Rausch SK, Shamansky L, Treat ML, Ziegler MM (1984) Cloning of the luciferase structural genes from Vibrio harveyi and expression of bioluminescence in Escherichia coli. Biochemistry 23:3663–3667

    Google Scholar 

  • Bergström S, Olsson O, Normark S (1982) Common evolutionary origin of chromosomal beta-lactamase genes in Enterobacteria. J Bacteriol 150:528–534

    Google Scholar 

  • Bertani G (1951) Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol 62:293–300

    Google Scholar 

  • Bolivar F, Rodriguez RL, Green PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S (1977) Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113

    Google Scholar 

  • Boyer MW, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in E. coli. J Mol Biol 41:121–136

    Google Scholar 

  • Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–251

    Google Scholar 

  • Carmi OA, Stewart GSAB, Wlitzur S, Kuhn J (1987) Use of bacterial luciferase to establish a promoter probe vehicle capable of nondestructive real-time analysis of gene expression in Bacillus spp. J Bacteriol 169:2165–2170

    Google Scholar 

  • Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the p15A cryptic miniplasmid. J Bacteriol 134:1141–1156

    Google Scholar 

  • Cohn DH, Milcham AJ, Simon MI, Nealson KH (1985) Nucleotide sequence of the luxA gene of Vibrio harveyi and the complete amino acid sequence of the α subunit of bacterial luciferase. J Biol Chem 260:6139–6146

    Google Scholar 

  • Engebrecht J, Simon M, Silverman M (1985) Measuring gene expression with light. Science 227:1345–1347

    Google Scholar 

  • Gold L, Pribnow D, Schneider T, Shinedling S, Swebelius-Singer B, Stormo G (1981) Translational initiation in procaryotes. Ann Rev Microbiol 35:365–403

    Google Scholar 

  • Jefferson RA, Burgess SM, Hirsh D (1986) β-Glucuronidase from Escherichia coli as a gene-fusion marker. Proc Natl Acad Sci USA 83:8447–8451

    Google Scholar 

  • Johnston TC, Thompson RB, Baldwin TO (1986) Nucleotide sequence of the luxB gene of Vibrio harveyi and the complete amino acid sequence of the β subunit of bacterial luciferase. J Biol Chem 261:4805–4811

    Google Scholar 

  • Koncz C, Olsson O, Langridge WHR, Schell J, Szalay AA (1987) Expression and assembly of functional bacterial luciferase in plants. Proc Natl Acad Sci USA 84:131–135

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Legocki RP, Legocki M, Baldwin TO, Szalay AA (1986) Bioluminescence in soybean root nodules: Demonstration of a general approach to assay gene expression in vivo by using bacterial luciferase. Proc Natl Acad Sci USA 83:9080–9084

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) In: Molecular Cloning. A laboratory manual. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, New York

    Google Scholar 

  • Meighen EA, Bartlet I (1980) Complementation of subunits from different bacterial luciferases. Evidence for the role of the β subunit in the bioluminescent mechanism. J Biol Chem 255:11181–11187

    Google Scholar 

  • Meighen EA, Nicoli MZ, Hastings JW (1971 a) Hybridization of bacterial luciferase with a variant introduced by chemical modification. Biochemistry 10:4062–4068

    Google Scholar 

  • Meighen EA, Nicoli MZ, Hastings JW (1971 b) Functional differences of the non-identical subunits of bacterial luciferase. Properties of hybrids of native and chemically modified bacterial luciferase. Biochemistry 10:4069–4073

    Google Scholar 

  • Nicoli MZ, Meighen EA, Hastings JW (1974) Bacterial luciferase: Chemistry of reactive sulfhydryl. J Biol Chem 249:2385–2392

    Google Scholar 

  • Norlander J, Kempe T, Messing J (1983) Construction of improved M13 vectors using oligodeoxynucleotide directed mutagenesis. Gene 26:101–106

    Google Scholar 

  • Ow DW, Wood KW, DeLuca M, DeWet JR, Helinski DR, Howell SH (1986) Transient and stable expression of the firefly luciferase gene in plant cells and transgenic plants. Science 234:856–859

    Google Scholar 

  • Rosenberg AH, Lade BN, Chui D, Dunn JJ, Studier FW (1987) Vectors for selective expression of cloned DNAs by T7 RNA polymerase. Gene 56:125–135

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Schmetterer G, Wolk CP, Elhay J (1986) Expression of luciferases from Vibrio harveyi and Vibrio fischeri in filamentous cyanobacteria. J Bacteriol 167:411–414

    Google Scholar 

  • Shine J, Dalgarno L (1974) The 3′-terminal sequence of Escherichia coli 16S ribosomal RNA: Complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346

    Google Scholar 

  • Studier FW, Moffatt BA (1986) Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol 189:113–130

    Google Scholar 

  • Tobin H, Stahelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    Google Scholar 

  • Ziegler MM, Baldwin TO (1981) Biochemistry of bacterial bioluminescence. Curr Top Bioenerg 12:65–113

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by W. Goebel

Rights and permissions

Reprints and permissions

About this article

Cite this article

Olsson, O., Koncz, C. & Szalay, A.A. The use of the luxA gene of the bacterial luciferase operon as a reporter gene. Mol Gen Genet 215, 1–9 (1988). https://doi.org/10.1007/BF00331295

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00331295

Key words

Navigation