- Email: [email protected]
Sequence of the goat αs2-casein-encoding cDNA
Gette, 125 (1993) 235-236 $j 1993 Elsevicr Science Publishers
GENE
B.V. AL1 rights reserved.
235
0378-I 119~93~S~6.~
06984
Sequence of the goat a,,-casein-encoding
cDNA*
(Mammary gland; lactoprotein; caprine gene)
Christine
Bouniol
Received by G. Bernardi:
24 September
1992; Revised/Accepted:
--.----
26 November/30
--.”
..,__-
November
1992; Received at pubfishers:
--
10 December
1992
-
._I
SUMMARY
The complete nucleotide sequence of a caprine ~1,~ -casein-encoding &DNA and the deduced 223-amino-acid sequence of pre-isl,,-casein were determined.
The a,,-casein (cc,,-Cas) is the most phospharylated Cas with up to 13 phosphate groups in bovine species (Brignon et al., 1977). The N- and C-terminal halves of the bovine mature Cas share 40% sequence identity, which suggests that the ancestral gene underwent an
internal duplication (Brignon et al., 1977). The sequences of bovine (Stewart et al., 1987), ovine (Boisnard and Petrissant, 1985), rat (Hobbs and Rosen, 1982), guineapig (Hall et al., 1984), mouse (Hennighausen et al., 1982) and pig (Alexander et al., 1992) cl&as cDNAs are
Fig. I. The nt sequence of a cDNA clone encoding the cap&e cx,,-Cas and the deduced aa sequence. A lactating caprine mammary gland cDNA library constructed in pUCl8 (SmaI site) was probed with an ovine a&as cDNA (Boisnard et Pet&ant, 1985). The positive clones were sequenced on both strands (Sanger et al., 1977) after subcloning. The nt sequence (upper line) and the deduced 208 aa sequence (lower line) are given. The stop codon is marked with three asterisks. Similar nt sequence repeats are underlined. Potential phosphorylated aa (Ser, Thr) are underlined with dashed lines. The putative polyadenylation site is doubly underlined. The 15-aa signal peptide is in bold-face type. GenBank accession No. X65160. Correspondence to: Dr. C. Houniol, Laboratoire de Genetique Biochimique, Institut National de la Recherche Agronomique, France. Tel. (33-l) 34-65-Z-75; Fax (33-I) 34-65-22-73. *On request. the author will supply detailed experimental evidence for the conclusions reached in this Brief Note. Abbreviations:
aa, amino
acid(s); Cas, casein; cDNA,
DNA complementary
to RNA; nt, nucieotidefs).
78350 Jouy-en-Josas,
236 known. Analysis of an a,,-Cas-encoding clone isolated from a caprine mammary gland cDNA library and 5’ primer extension (6 nt) gave the complete sequence of the 1013 nt aSZ-CasA cDNA (Fig. 1). The 669-nt coding frame, which contains two similar (60%) sequences of 222 nt, is flanked by S- and 3’-untranslated regions of 42 and 302 nt, respectively. This cDNA shares 98.5%, 94.5% and 75.3% similarity with ovine, bovine and porcine a,*-Cas cDNAs but is quite different from its shorter rodent counterparts, in particular that of the mouse (about 700 nt) which lacks the duplicated sequence.
ACKNOWLEDGEMENTS
REFERENCES Alexander, L.J., Das Gupta, N.A. and Beattie, C.W.: The sequence of porcine a,,-casein cDNA. Anim. Genet. 23 (1992) 365-367. Boisnard, M. and Pitrissant, G.: Complete sequence of ovine cc,,-casein messenger RNA. Biochimie 67 (1985) 1043-1051, Brignon, G., Ribadeau-Dumas, B., Mercier, J.C., Pehssier J.P. and Das, B.C.: Complete amino-acid sequence of bovine a,,-casein. FEBS Lett. 76 (1977) 274-279. Hall, L., Laird, J.E., Pascall, J.C. and Craig, R.K.: Guinea-pig casein A cDNA. Nucleotide sequence analysis and comparison of the deduced protein sequence with that of bovine r,,-casein. Eur. J. Biothem. i 38 (1984) 585-589. Hennighausen, L.G_ Steudle, A. and Sippel. A.E.: Nucleotide sequence of cloned cDNA coding for mouse &-casein. Eur. J. Biochem. 126 (1982) 569-572. Hobbs, A.A. and Rosen J.M.: Sequence of rat TX-and y-casein mRNAs: evolutionary comparison of the calcium-dependent rat casein multigene family. Nucleic Acids Res. 10 (1982) 807998098. Sanger, F., Nicklen, S. and Coulson, A.R.: DNA sequencing with chainterminating
I thank Dr. C. Leroux for the construction of the cDNA library. Many thanks to C. Printz and J.C. Mercier for valuable discussions and critical reading of the manuscript.
inhibitors.
Proc.
Natl.
Acad.
Sci.
USA
74 (1977)
5463-5467. Stewart, A.F., Bonsing, J., Beattie, C.W., Shah, F., Willis, 1.M. and Mackinlay, A.G.: Complete nucleotide sequence of bovine otsz- and R-casein cDNAs: comparisons with related other sequences in other species. Mol. Biol. Evol. 4 (1987)23 l-241.
GENE
B.V. AL1 rights reserved.
235
0378-I 119~93~S~6.~
06984
Sequence of the goat a,,-casein-encoding
cDNA*
(Mammary gland; lactoprotein; caprine gene)
Christine
Bouniol
Received by G. Bernardi:
24 September
1992; Revised/Accepted:
--.----
26 November/30
--.”
..,__-
November
1992; Received at pubfishers:
--
10 December
1992
-
._I
SUMMARY
The complete nucleotide sequence of a caprine ~1,~ -casein-encoding &DNA and the deduced 223-amino-acid sequence of pre-isl,,-casein were determined.
The a,,-casein (cc,,-Cas) is the most phospharylated Cas with up to 13 phosphate groups in bovine species (Brignon et al., 1977). The N- and C-terminal halves of the bovine mature Cas share 40% sequence identity, which suggests that the ancestral gene underwent an
internal duplication (Brignon et al., 1977). The sequences of bovine (Stewart et al., 1987), ovine (Boisnard and Petrissant, 1985), rat (Hobbs and Rosen, 1982), guineapig (Hall et al., 1984), mouse (Hennighausen et al., 1982) and pig (Alexander et al., 1992) cl&as cDNAs are
Fig. I. The nt sequence of a cDNA clone encoding the cap&e cx,,-Cas and the deduced aa sequence. A lactating caprine mammary gland cDNA library constructed in pUCl8 (SmaI site) was probed with an ovine a&as cDNA (Boisnard et Pet&ant, 1985). The positive clones were sequenced on both strands (Sanger et al., 1977) after subcloning. The nt sequence (upper line) and the deduced 208 aa sequence (lower line) are given. The stop codon is marked with three asterisks. Similar nt sequence repeats are underlined. Potential phosphorylated aa (Ser, Thr) are underlined with dashed lines. The putative polyadenylation site is doubly underlined. The 15-aa signal peptide is in bold-face type. GenBank accession No. X65160. Correspondence to: Dr. C. Houniol, Laboratoire de Genetique Biochimique, Institut National de la Recherche Agronomique, France. Tel. (33-l) 34-65-Z-75; Fax (33-I) 34-65-22-73. *On request. the author will supply detailed experimental evidence for the conclusions reached in this Brief Note. Abbreviations:
aa, amino
acid(s); Cas, casein; cDNA,
DNA complementary
to RNA; nt, nucieotidefs).
78350 Jouy-en-Josas,
236 known. Analysis of an a,,-Cas-encoding clone isolated from a caprine mammary gland cDNA library and 5’ primer extension (6 nt) gave the complete sequence of the 1013 nt aSZ-CasA cDNA (Fig. 1). The 669-nt coding frame, which contains two similar (60%) sequences of 222 nt, is flanked by S- and 3’-untranslated regions of 42 and 302 nt, respectively. This cDNA shares 98.5%, 94.5% and 75.3% similarity with ovine, bovine and porcine a,*-Cas cDNAs but is quite different from its shorter rodent counterparts, in particular that of the mouse (about 700 nt) which lacks the duplicated sequence.
ACKNOWLEDGEMENTS
REFERENCES Alexander, L.J., Das Gupta, N.A. and Beattie, C.W.: The sequence of porcine a,,-casein cDNA. Anim. Genet. 23 (1992) 365-367. Boisnard, M. and Pitrissant, G.: Complete sequence of ovine cc,,-casein messenger RNA. Biochimie 67 (1985) 1043-1051, Brignon, G., Ribadeau-Dumas, B., Mercier, J.C., Pehssier J.P. and Das, B.C.: Complete amino-acid sequence of bovine a,,-casein. FEBS Lett. 76 (1977) 274-279. Hall, L., Laird, J.E., Pascall, J.C. and Craig, R.K.: Guinea-pig casein A cDNA. Nucleotide sequence analysis and comparison of the deduced protein sequence with that of bovine r,,-casein. Eur. J. Biothem. i 38 (1984) 585-589. Hennighausen, L.G_ Steudle, A. and Sippel. A.E.: Nucleotide sequence of cloned cDNA coding for mouse &-casein. Eur. J. Biochem. 126 (1982) 569-572. Hobbs, A.A. and Rosen J.M.: Sequence of rat TX-and y-casein mRNAs: evolutionary comparison of the calcium-dependent rat casein multigene family. Nucleic Acids Res. 10 (1982) 807998098. Sanger, F., Nicklen, S. and Coulson, A.R.: DNA sequencing with chainterminating
I thank Dr. C. Leroux for the construction of the cDNA library. Many thanks to C. Printz and J.C. Mercier for valuable discussions and critical reading of the manuscript.
inhibitors.
Proc.
Natl.
Acad.
Sci.
USA
74 (1977)
5463-5467. Stewart, A.F., Bonsing, J., Beattie, C.W., Shah, F., Willis, 1.M. and Mackinlay, A.G.: Complete nucleotide sequence of bovine otsz- and R-casein cDNAs: comparisons with related other sequences in other species. Mol. Biol. Evol. 4 (1987)23 l-241.