Nucleic Acids Research, Vol. 18, No. 23 7175

The molecular cloning of ovine interleukin 2 polymerase chain reaction

gene

by the

H.-F.Seow, J.S.Rothel, A.J.Radford and P.R.Wood CSIRO, Division of Animal Health, Animal Health Research Laboratory, Private Bag No. 1, Parkville, Victoria 3052, Australia Submitted October 19, 1990

Interleukin 2 (IL-2), formerly known as T-cell growth factor, is a lymphokine produced by mitogen-activated T cells (1, 2). IL-2 regulates the growth and differentiation of lymphopoietic cells; for example, it can cause maturation of cytotoxic T-cells, induce 'y-interferon production and promote natural killer and lymphokine-activated killer cell activity. A cDNA clone of human IL-2 was initially isolated by Taniguchi et al. (3). Murine and bovine genes have also been cloned (4, 5). We now report the cloning of the ovine IL-2 gene using the polymerase chain reaction with primers based on the bovine IL-2 sequence at the 5' and 3' ends of the gene. The sequence of the primers were ATG TAC AAG ATA CAA CTC TTG TCT T and GTC ATT GTT GAG TAG ATG CTT TGA C. Complementary DNA was synthesized using avian myeloblastosis virus reverse transcriptase of RNA isolated from 24 h Concanavalin A (7.5 pg/ml) and phorbol-myristate acetate (7.5 ng/ml) stimulated lymph node cells. The amplified fragment was cloned into a plasmid vector pUC 19 and the DNA sequence was determined by the chain termination method. The ovine

EMBL accession no. X55641

sequence shows an overall homology of 98% with the bovine sequence at nucleotide level and 97% at amino acid level.

ACKNOWLEDGEMENTS We wish to thank Marie-Jean David for technical support and Leigh Corner for professional assistance.

REFERENCES 1. Morgan,D.A., Ruscetti,F.W. and Gallo,R. (1976) Science (Washington D. C.) 193, 1007-1008. 2. Gilfis,S., Ferm,M.M., Ou,W. and Smith,K.A. (1978) J. Immunol. 120, 2027-2032. 3. Taniguchi,T., Matsui,H., Fujita,T., Takaoka,C., Kashima,N., Yoshimoto,R. and Hamuro,J. (1983) Nature (Lond.) 302, 305-310. 4. Kashima,N., Nishi-Takaoka,C., Fujita,T., Taki,S., Yamada,G., Hamuro,J. and Taniguchi,T. (1985) Nature (Lond.) 313, 402-404. 5. Cerretti,D.P., McKereghan,K., Larsen,A., Cantrell,M.A., Anderson,D., Gillis,S., Cosman,D. and Baker,P.E. (1986) Proc. Natl. Acad. Sci. USA 83, 3223-3227.

1 1

Met Tyr Lys Ile Gln Leu Leu Ser Cys Ile Ala Leu Thr Leu Ala Leu Val Ala Asn Gly Ala Pro Thr Ser ATG TAC AAG ATA CAA CTC TTG TCT TGC ATT GCA CTA ACT CTT GCA CTC GTT GCA AAC GGT GCA CCT ACT TCA

24 72

25 73

Ser Ser Thr Gly Asn Thr Met Lys Glu Val Lys Ser Leu Leu Leu Asp Leu Gln Leu Leu Leu Glu Lys Val AGC TCT ACG GGG AAC ACA ATG AAA GAA GTG AAG TCA TTG CTG CTG GAT TTA CAG TTG CTT TTG GAG AAA GTT

48 144

49 145

Lys Asn Pro Glu Asn Leu Lys Leu Ser Arg Met His Thr Phe Asn Phe Tyr Met Pro Lys Val Asn Ala Thr AAA AAT CCC GAG AAC CTC AAG CTC TCC AGG ATG CAT ACA TTT AAC TTC TAC ATG CCC AAG GTT AAC GCT ACA

72 216

73 217

Glu Leu Lys His Leu Lys Cys Leu Leu Glu Glu Leu Lys Leu Leu Glu Glu Val Leu Asp Leu Ala Pro Ser GAA TTG AAA CAT CTT AAG TGT TTA CTA GAA GAA CTC AAA CTT CTA GAG GAA GTG CTA GAT TTA GCT CCA AGC

96 288

97 289

Lys Asn Leu Asn Thr Arg Glu Ile Lys Asp Ser Met Asp Asn Ile Lys Arg Ile Val Leu Glu Leu Gln Gly AAA AAC CTG AAC ACC AGA GAG ATC AAG GAT TCA ATG GAC AAT ATC AAG AGA ATA GTT TTG GAA CTA CAG GGA

120 360

121 361

Ser Glu Thr Arg Phe Thr Cys Glu Tyr Asp Asp Ala Thr Val Lys Ala Val Glu Phe Leu Asn Lys Trp Ile TCT GAA ACA AGA TTC ACA TGT GAA TAT GAT GAT GCG ACA GTA AAG GCT GTA GAA TTT CTG AAC AAA TGG ATT

144 432

145 433

Thr Phe Cys Gln Ser Ile Tyr Ser Thr Met Thr *** ACC TTT TGT CAA AGC ATC TAC TCA ACA ATG ACT TGA

The molecular cloning of ovine interleukin 2 gene by the polymerase chain reaction.

Nucleic Acids Research, Vol. 18, No. 23 7175 The molecular cloning of ovine interleukin 2 polymerase chain reaction gene by the H.-F.Seow, J.S.Rot...
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