Cloning, sequencing, and characterization of CYP1A1 cDNA from leaping mullet (Liza Saliens) liver and implications for the potential functions of its conserved amino acids
✍ Scribed by Alaattin Sen; Chin-Hwa Hu; Ena Urbach; Jun-Lan Wang-Buhler; Yea-Huey Yang; Emel Arinc; Donald R. Buhler
- Publisher
- John Wiley and Sons
- Year
- 2001
- Tongue
- English
- Weight
- 862 KB
- Volume
- 15
- Category
- Article
- ISSN
- 1095-6670
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✦ Synopsis
Abstract
A 2,037 bp CYP1A1 cDNA (GenBank AF072899) was cloned through screening of a λZipLox cDNA library constructed from the liver of a leaping mullet (Liza saliens) fish captured from Izmir Bay on the Aegean coast of Turkey using rainbow trout CYP1A1 cDNA as a probe. This clone has a 130 bp 5'‐flanking region, a 1,563 bp open reading frame (ORF) encoding a 521‐amino acid protein (58,972 Da), and a 344 bp 3'‐untranslated region without a poly (A) tail. Alignment of the deduced amino acids of CYP1A1 cDNAs showed 58% and 69–96% identities with human and 12 other fish species, respectively. Southern blot analysis suggested that this CYP1A1 cDNA was from a single‐copy gene. Based on the comparison with CYP1A1 genes reported for fish and mammals, the leaping mullet CYP1A1 gene is probably split into 7 exons. The intron insertion sites were predicted. Alignment of the CYP1A1 cDNA encoded amino acids from 13 fish and 7 mammalian species disclosed differences in highly conserved amino acids between aquatic and land vertebrates. The possible associated secondary structure; conserved motifs and substrate‐binding sites were discussed. The phylogenetic relationships of CYP1A1s among 13 fish species were analyzed by a distance method. © 2001 John Wiley & Sons, Inc. J Biochem Mol Toxicol 15:243–255, 2001