Cdna-derived Amino-acid Sequence Of A Land Turtle (geochelone Carbonaria) β-chain Hemoglobin

The cDNA sequence encoding the turtle Geochelone carbonaria β-chain was determinated. The isolation of hemoglobin mRNA was based on degenerate primers' PCR in combination with 5'- and 3'-RACE protocol. The full length cDNA is 615 bp with the ATG start codon at position 53 and TGA stop codon at position 495; The AATAAA polyadenylation signal is found at position 599. The deduced polypeptyde contains 146 amino-acid residues. The predicted amino acid sequence shares 83% identity with the β-globin of a related specie, the aquatic turtle C. p. belli. Otherwise, identity is higher when compared with chicken β-Hb (80%) than with other reptilian orders (Squamata, 69%, and Crocodilia, 61%). Compared with human HbA, there is 67% identity, and at least three amino acid substitutions could be of some functional significance (Glu43β→Ser, His116β→Thr and His143β→Leu). To our knowledge this represents the first cDNA sequence of a reptile globin gene described.422255260Komiyama, N.H., Miyazaki, G., Tame, J., Nagai, K., (1995) Nature, 373, pp. 244-246Mylvaganam, S.E., Bonaventura, C., Bonaventura, J., Getzoff, E.D., (1996) Nature Struct. Biol., 3, pp. 275-283Brittain, T., (1991) Comp. Biochem. Physiol., 99 B, pp. 731-740Rough, F.H., (1980) Am. Zool., 20, pp. 173-184Torsoni, M.A., Viana, R.I., Stoppa, G.R., Barros, B.F., Cesquini, M., Ogo, S.H., J. Comp. Biochem. Physiol., , submittedRücknagel, K.P., Reischl, E., Braunitzer, G., (1984) Hoppe-Seyler's Z. Physiol. Chem. B, 365, pp. 1163-1171Torsoni, M.A., Souza, A.N., Ogo, S.H., J. Comp. Physiol., , submittedChomczynski, P., Sacchi, N., (1987) Analyt. Biochem., 162, pp. 156-159Patient, R.K., Harris, R., Walmsley, M.E., Williams, J.G., (1983) J. Biol. Chem., 258, pp. 8521-8523Richards, R.I., Shine, J., Ullrich, A., Wells, J.R.E., Goodman, H.M., (1979) Nucl. Ac. Res., 7, pp. 1137-1146D'Avino, R., Caruso, C., Tamburrini, M., Romano, M., Rutigliano, B., De Laureto, P.P., Camardella, L., Di Prisco, G., (1994) J. Biol. Chem., 269, pp. 9675-9681Lawn, R.M., Efstratiadis, A., O'Connell, C., Maniatis, T., (1980) Cell, 21, pp. 647-651Frohman, M.A., (1990) PCR Protocols. A Guide to Methods and Applications, pp. 28-38. , Ed. Innis, M.A., Gelfand, D.H., Sninsky, J.J. and White, T.J., Academic Press, CaliforniaRücknagel, K.P., Braunitzer, G., (1988) Biol. Chem. Hoppe-Seyler, 329, pp. 123-131Arnone, A., (1972) Nature, 237, pp. 146-148Leclercq, F., Schnek, A.G., Braunitzer, G., Stangl, A., Schrank, B., (1981) Hoppe-Seyler's Physiol. Chem., 362, pp. 1151-1158Matsuura, M.S.A., Fushitani, K., Riggs, A.F., (1989) J. Biol. Chem, 264, pp. 5515-552