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Rapid Communication: Nucleotide Sequence of Rainbow Trout a-Globin I and IV cDNA Goro Yoshizaki, Atsuko Takano, Takashi Aoki, and Fumio Takashima Department of Aquatic Biosciences, Tokyo University of Fisheries, Konan, Minato-ku, Tokyo 108, Japan Name of Sequence. Rainbow trout a-globin I and IV cDNA. EMBL/GenBank Accession Number. a-globin I, D88113; a-globin IV, D88114. Species. Oncorhynchus mykiss. Origin of Clone. Messenger RNA was isolated from total blood cells. Complementary DNA was synthesized using the cDNA Synthesis Kit (Pharmacia Biotech, Uppsala, Sweden). A library was then constructed by cloning cDNA into pUC118. The library was screened with carp a-globin cDNA (Takeshita et al., 1984). Two positive clones (clone 1 and 5 ) were identified and sequenced with the dideoxy method. Comparison with Related Sequence. The deduced amino acid sequence of clone 1 showed 97.9% identity to rainbow trout a-globin IV (Bossa et al., 1983). Clone 5 showed 97.9% identity to rainbow trout aglobin I (Petruzzelli et al., 1989). Clones 1 and 5 demonstrated 61.3% amino acid identity with one another. Also, the amino acid identities of clone 1 and 5 to human a-globin (Kleinschmidt and Sgouros, 1987) were 50.4% and 56.7%, respectively. Sequence Data. The nucleotide and deduced amino acid sequence of a-globin I and IV are shown in Figures 1A and B, respectively. These clones had an open reading frame that encoded 143 amino acids. Comments. The identity data indicate that clones 1 and 5 encode a-globin IV and I, respectively. However, the deduced amino acid sequence of clone 5 was two residues longer than that of a-globin I. The distal and proximal histidines (Figure 1; double underline) were identified, and the predicted heme contact region (Figure 1; underline) was found to be well conserved (i.e., approximately 85% identity) between the rainbow trout and human sequences. (Salmo irideus). Biochim. Biophys. Acta 995:255. Takeshita, S., T. Aoki, Y. Fukumaki, and Y. Takagi. 1984. Cloning and sequence analysis of a cDNA for the a-globin mRNA of carp, Cyprinus carpio. Biochim. Biophys. Acta 783:265. Key Words: Rainbow Trout, Complementary DNA, Nucleotide Sequences, Cloning, Amino Acid Sequences J. Anim. Sci. 1997. 75:1426 Literature Cited Bossa, F., D. Barra, R. Petruzzelli, F. Martini, and M. Brunori. 1983. Primary structure of hemoglobin from trout ( Salmo irideus) . Amino acid sequence of a-chain of trout Hb I. Biochim. Biophys. Acta 536:298. Kleinschmidt, T., and J. G. Sgouros. 1987. Hemoglobin sequences. Biol. Chem. Hoppe-Seyler 368:579. Petruzzelli, R., D. Barra, L. Sensi, F. Bossa, and M. Brunori. 1989. Amino acid sequence of a-chain of hemoglobin IV from trout Received October 7, 1996. Accepted January 16, 1997. Figure 1. Nucleotide and deduced amino acid sequences of rainbow trout a-globin I (A) and IV (B). Nucleotide number and the amino acid residue number are shown at right. Distal (amino acid number 60) and proximal histidines (amino acid number 89) are doubleunderlined and the heme-contact regions are underlined. 1426