Isolation and expression of a cDNA encoding Renilla reniformis luciferase.
Renilla reniformis is an anthozoan coelenterate capable of exhibiting bioluminescence. Bioluminescence in Renilla results from the oxidation of coelenterate luciferin (coelenterazine) by luciferase [Renilla-luciferin:oxygen 2-oxidoreductase (decarboxylating), EC 1.13.12.5]. In vivo, the excited state luciferin-luciferase complex undergoes the process of nonradiative energy transfer to an accessory protein, green fluorescent protein, which results in green bioluminescence. In vitro, Renilla luciferase emits blue light in the absence of any green fluorescent protein. A Renilla cDNA library has been constructed in lambda gt11 and screened by plaque hybridization with two oligonucleotide probes. We report here the isolation and characterization of a luciferase cDNA and its gene product. The recombinant luciferase expressed in Escherichia coli is identical to native luciferase as determined by SDS/PAGE, immunoblot analysis, and bioluminescence emission characteristics.
Lorenz
W W
WW
Department of Biochemistry, University of Georgia, Athens 30602.
McCann
R O
RO
Longiaru
M
M
Cormier
M J
MJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003063:Cnidaria / Q000235:genetics*; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D015245:Deoxyribonuclease BamHI; D015246:Deoxyribonuclease EcoRI; D004735:Energy Transfer; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D049454:Firefly Luciferin / Q000378:metabolism; D015870:Gene Expression; D008156:Luciferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D013053:Spectrophotometry; D014169:Transformation, Bacterial
D015345:Oligonucleotide Probes; D049454:Firefly Luciferin; D004247:DNA; D008156:Luciferases; D015245:Deoxyribonuclease BamHI; D015246:Deoxyribonuclease EcoRI