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Complementary DNA coding click beetle luciferases can elicit bioluminescence of different colors
K V Wood1, Y A Lam, H H Seliger
1Department of Chemistry, University of California, San Diego, La Jolla 92093.
Science (New York, N.Y.)
|May 12, 1989
Summary
Researchers isolated click beetle complementary DNA (cDNA) clones that produce visible bioluminescence in bacteria. These clones code for four distinct luciferases, useful as multiple reporter genes in molecular biology experiments.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- Bioluminescence is a natural light-emitting process found in various organisms.
- Luciferases are enzymes that catalyze bioluminescence reactions.
- Click beetles (Pyrophorus plagiophthalamus) possess unique bioluminescent properties.
Purpose of the Study:
- To isolate and characterize complementary DNA (cDNA) clones encoding luciferases from the click beetle, Pyrophorus plagiophthalamus.
- To evaluate the potential of these clones as reporter genes in molecular biology.
Main Methods:
- Messenger RNA (mRNA) was isolated from the light organs of Pyrophorus plagiophthalamus.
- Complementary DNA (cDNA) libraries were constructed and eleven clones were sequenced.
- The isolated cDNA clones were expressed in Escherichia coli to assess their bioluminescent capabilities.
Main Results:
- Eleven cDNA clones were successfully generated, capable of eliciting visible bioluminescence when expressed in E. coli.
- The clones encode four distinct types of luciferases, producing green, yellow-green, yellow, and orange light (546-593 nm).
- These beetle luciferases share 95-99% amino acid sequence identity with each other but only 48% with firefly luciferase.
Conclusions:
- The characterized click beetle luciferases represent novel tools for bioluminescence applications.
- The distinct colors emitted by these luciferases make them suitable for multiplexing in reporter gene assays.
- These findings expand the toolkit for researchers utilizing bioluminescent reporters in various biological studies.
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