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Nucleic Acids Research, 1982, Vol. 10, No. 7 2177-2187
© 1982

MOLECULAR BIOLOGY

Molecular cloning and nucleotide sequence of cDNA coding for calf preprochymosin

T.J.R. Harris*, P.A. Lowe*, A. Lyons*, P.G. Thomas*, M.A.W. Eaton+, T.A. Millican+, T.P. Patel+, C.C. Bose+, N.H. Carey** and M.T. Doel*

*Department of Molecular Biology Celltech Ltd., 250 Bath Road, Slough SL1 4DY, Berkshire, UK +Department of Chemistry Celltech Ltd., 250 Bath Road, Slough SL1 4DY, Berkshire, UK

Received March 8, 1982. Accepted March 25, 1982.

DNA complementary to calf stomach mRNA has been synthesised and inserted into the Pstl site of pAT153 by G-C tailing. Clones containing sequences coding for prochymesin were recognised by colony hybridisation with cDNA extended from a chemically synthesised oligodeoxynucleotide primer, the sequence of which was predicted from the published amino acid sequence of calf prochymesin1. Two clones were identified which together contained a complete copy of prochymesin mRNA. The nucleotide sequence is in substantial agreement with the reported amino acid sequence of prochymosin2 and shows that this protein has a mol.wt. of 40431 and chyemsin a mol.wt. of 35612. The sequence also indicates that prochyn is synthesised as a precursor melecule, preprochymosin, having a 16 amino acid hydrophobic leader sequence analogous to that reported for other secreted proteins3.

**Research Director.


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