Interleukin 2 (IL-2) is a lymphokine produced by activatedT lymphocytes. ... murine (9) and bovine (10, 11) IL-2 have been cloned, sequenced and expressed ...
Nucleic Acids Research, Vol. 18, No. 19 5883
k.-1/ 1990 Oxford University Press
cDNA cloning
ovine interleukin 2
of
PCR
by
Jane C.Goodall1, David C.Emery, Anthony C.F.Perry, Leonard S.Englishl and Len Hall* Department of Biochemistry, University of Bristol, Bristol BS8 1TD and 'Department of Science, Bristol Polytechnic, Bristol BS16 1QY, UK EMBL accession
Submitted August 28, 1990
Morgan,D.A. et al. (1976) Science 193, 1007-1008. Robb,R.J. et al. (1981) J. Exp. Med. 154, 1455-1474. Smith,K.A. (1988) Science 240, 1169-1176. Waldmann,T.A. et al. (1984) J. Exp. Med. 160, 1450-1466. Boyd,A.W. et al. (1985) J. Immunol. 134, 2387-2392. Siegel,J.P. et al. (1987) Science 238, 75-78. Devos,R. et al. (1983) Nucl. Acids Res. 11, 4307-4323. Taniguchi,T. et al. (1983) Nature 303, 305-310. Kashima,N. et al. (1985) Nature 313, 402-405. Cerretti,D.P. et al. (1986) Proc. Natl. Acad. Sci. USA 83, 3223-3227. Reeves,R. et al. (1986) Proc. Natl. Acad. Sci. USA 83, 3228-3232. English,L.S. et al. (1985) Cell. Imnunnol. 90, 314-321. English,L.S. et al. (1985) Vet. Immunol. Immunopathol. 9, 59-69.
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13.
ATG TAC MAG ATA CAA CCC TTG TCT TGC ATT GCA CTA ACT CTT GCA CTC GTT GCA MC GGT GCA CCT ACT TCA AGC TCT ACG Q G A P T S S S T A T L L V A N A L I M Y K I P L S C -1 +1 -10 -20 (L)
82
GGG
MC
G
N
M
K
GM
GTG
E
V
AAG K
TCA
TTG
CTG
CTA
S
L
L
L
GAT
D 20
TTA
CAG
TTG
CTT
TTG
GAG
AM
GTT
L
Q
L
L
L
E
K
V
AMA AAT N K 30
CCC
GAG
AAC
CTC
P
E
N
L
163
MAG CTC TCC AGG ATG CAT ACA TTT MC TTC TAC ATG CCC AAG GTT AAC GCT ACA GAA TTG AAA CAT CTT AAG TGT TTA CTA L K C L H L K E L F Y M P K V N A T F N K L S R M H T 60 50 40 (V) (D)
244
GAA GAA CTC AM CTT CTA GAG GAA GTG CTA GAT TTA GCT CCA AGC AAA AAC CTG AAC ACC AGA GAG ATC I E T R N D N L P S K L A E L K L L E L E E V 80 (P) 70 (N)
AAG K
325
GAC AAT ATC AAG AGA ATA GTT TTG GAA CTA CAG GGA TCT GAA ACA AGA TTC ACA TGT GAA TAT GAT GAT D D C E T Y F R K R I V L E L Q G S E T D N I 110 100 90
GCG ACA GTA AAG K V A T (N)
406
GCT GTA GAA TTT CTG AAC AAA TGG ATT ACC TTT TGT CAA AGC ATC TAC TCA ACA ATG T I M W Y S T F C Q S A V E F N K I L 120
492
*
REFERENCES
1
T 10
130
ACT TGA T * 135
ACTATCAGGCTTTCTATTTATTTAMATATTTAAAATTTATATTTATTTTTTGATATATGTTTTCCTACCTTTTGTAACTATTAGT
To whom correspondence should be addressed
X53934
responsible for species specificity (12, 13). Future work will involve expressing the cloned ovine IL-2 cDNA for affinity labelling studies using human and sheep activated T cells.
Interleukin 2 (IL-2) is a lymphokine produced by activated T lymphocytes. It supports clonal expansion of helper and cytotoxic T cells (1-3), growth and differentiation of B cells (4, 5) and activates natural killer cells (6). The cDNAs of human (7, 8), murine (9) and bovine (10, 11) IL-2 have been cloned, sequenced and expressed (7-11). We have now cloned and sequenced the entire coding region of ovine IL-2 cDNA by PCR, using primers based on the bovine sequence (10, 11). The predicted precursor protein consists of 155 amino acids, representing a mature protein of 135 residues and a signal peptide of 20 residues. Comparison of human, murine, bovine and ovine IL-2 sequences reveals extensive regions of identity. However, the similarity of the bovine and ovine sequences is particularly striking, with only 16 nucleotide and 6 amino acid differences (the latter shown below in parentheses) within the analogous sequenced regions. Such comparisons should help to identify those residues in ovine IL-2
ACA ATG AAA
no.
GAT D
TCA ATG S M
TAACTGTGTGCTTCTCATTTTAA
