Cloning of the Vibrio harveyi luciferase genes: use of a synthetic oligonucleotide probe

Creators: Cohn, Daniel H.; Ogden, Richard C.; Abelson, John N.; Baldwin, Thomas O.; Nealson, Kenneth H.; Simon, Melvin I.; Mileham, Alan J.

Abstract

A mixed-sequence synthetic oligonucleotide probe was used to isolate a clone containing the gene encoding the alpha subunit of bacterial luciferase from Vibrio harveyi and part of the gene coding for the beta subunit. DNA sequence analysis has allowed us to determine that the genes are closely linked on the bacterial chromosome and transcribed in the same direction. Comparison of the sequences in the regions preceding the two structural genes has revealed considerable homology and has identified sites that may be involved in the expression of the genes. Identification of a clone from a clone bank of total genomic DNA from this organism shows that mixed probes can be successfully used to isolate a gene of interest from any bacterium provided some protein sequence for the gene product is available.

Additional Information

© 1983 by the National Academy of Sciences. Communicated by W.D. McElroy, October 7, 1982. We thank K. Itakura and S. Rausch, who provided unpublished results; A. Boyd for assistance with DNA sequence analyses; J. Engebrecht for technical assistance; and K. Janssen and R. Ng for critical reading of the manuscript. This work was supported by Office of Naval Research Grant N000-14-81-K-0343 and a National Science Foundation grant (PCM 82-41242) to T.O.B. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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