Molecular cloning and genomic organization of a gene for luciferin-binding protein from the dinoflagellate Gonyaulax polyedra

• Published in: The Journal of biological chemistry Vol. 268; no. 12; pp. 8842 - 8850
• Main Authors: DONG-HEE LEE, MITTAG, M, SCZEKAN, S, MORSE, D, HASTINGS, J. W
• Format: Journal Article
• Language: English
• Published: Bethesda, MD American Society for Biochemistry and Molecular Biology 25.04.1993
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; bioluminescence; Calcium-Binding Proteins - genetics; cDNA; Cloning, Molecular; Dinoflagellida - genetics; DNA; DNA, Protozoan; Fundamental and applied biological sciences. Psychology; genes; Genes, Protozoan; Genes. Genome; Gonyaulax polyedra; luciferin; luciferin-binding protein; luminous organisms; Marine; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; nucleotide sequence; predictions; proteins; Protozoan Proteins - genetics; Restriction Mapping; Binding protein; Gene; Complementary DNA; Gonyaulax polyedra; Luciferin; Algae; Pyrrophycophyta; Gene organization; Primary structure; Molecular cloning; Thallophyta
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)52950-1

The circadian expressed luciferin-binding protein (LBP) gene from the marine bioluminescent alga Gonyaulax polyedra represents the first dinoflagellate gene that has been cloned and sequenced at both cDNA and genomic levels. Starting with a fragment from the 3'-end of the LBP cDNA that was found by immunoscreening of a cDNA library, genomic clones were obtained by the inverse polymerase chain reaction technique. Full-length cDNA clones were selected by screening a cDNA library by plaque hybridizations and by polymerase chain reaction amplifications. The LBP sequence has a 2004-nucleotide open reading frame coding for a protein of 668 amino acids (approximately 75 kDa). The reading frame and identity of the clone were confirmed by the sequence of an octapeptide obtained from a purified fragment of CNBr-treated LBP. A variant LBP cDNA was found to differ in sequence by approximately 11% at the DNA level. The untranslated regions of the mRNA are 111 nucleotides (5'-untranslated region) and 158 nucleotides (3'-untranslated region) long, respectively. The LBP gene contains no introns and exhibits certain features not typical for a eukaryotic gene. Its promoter does not include the typical TATA box within approximately 50 nucleotides upstream of the transcription start site, and the usual poly(A+) signal (AAUAAA) is not present on the end of the LBP mRNA. The copy number of the gene is very high (approximately 1000 copies/cell). However, the universal genetic code and conserved positions relevant for the translational apparatus are maintained.