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Molecular cloning and nucleotide sequence of cDNA coding for calf preprochymosin.

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

DNA complementary to calf stomach mRNA has been synthesised and inserted into the Pst1 site of pAT153 by G-C tailing. Clones containing sequences coding for prochymosin 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 prochymosin. Two clones were identified which together contained a complete copy of prochymosin mRNA. The nucleotide sequence is in substantial agreement with the reported amino acid sequence of prochymosin and shows that this protein has a mol.wt. of 40431 and chymosin a mol.wt. of 35612. The sequence also indicates that prochymosin is synthesised as a precursor molecule, preprochymosin, having a 16 amino acid hydrophobic leader sequence analogous to that reported for other secreted proteins.

Nucleic Acids Res. 10:2177-2187(1982) [PubMed] [Europe PMC]

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