Nucleotide sequence of the luxA gene of Vibrio harveyi and the complete amino acid sequence of the alpha subunit of bacterial luciferase
The nucleotide sequence of the 1.85-kilobase EcoRI fragment from Vibrio harveyi that was cloned using a mixed-sequence synthetic oligonucleotide probe (Cohn, D. H., Ogden, R. C., Abelson, J. N., Baldwin, T. O., Nealson, K. H., Simon, M. I., and Mileham, A. J. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 120-123) has been determined. The alpha subunit-coding region (luxA) was found to begin at base number 707 and end at base number 1771. The alpha subunit has a calculated molecular weight of 40,108 and comprises a total of 355 amino acid residues. There are 34 base pairs separating the start of the alpha subunit structural gene and a 669-base open reading frame extending from the proximal EcoRI site. At the 3' end of the luxA coding region there are 26 bases between the end of the structural gene and the start of the luxB structural gene. Approximately two-thirds of the alpha subunit was sequenced by protein chemical techniques. The amino acid sequence implied by the DNA sequence, with few exceptions, confirmed the chemically determined sequence. Regions of the alpha subunit thought to comprise the active center were found to reside in two discrete and relatively basic regions, one from around residues 100-115 and the second from around residues 280-295.
Additional Information© 1985 by the American Society for Biochemistry and Molecular Biology. (Received for publication, October 15, 1984) We thank Dr. A. Boyd for assistance with DNA sequencing and secondary structure prediction. Dr. Rosemarie Swanson offered numerous insightful comments on the hydrophobicity considerations of the amino acid sequence, and Dr. Timothy Johnston provided the essence of our interpretation of the codon usage. This work was supported by grants from the Office of Naval Research (N000-14-81-K-0343 to M.I.S. and N000-14-800C-0066 to K.H.N.), and grants from the National Science Foundation (PCM 82-41242), the National Institutes of Health (AG-03697), the Robert A. Welch Foundation (A-865), and the United States Department of Agriculture (Hatch Grant RI 6545) to T.O.B. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Published - COHjbc85.pdf