Molecular cloning and nucleotide sequence of cDNA coding for calf preprochymosin

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...

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Bibliographic Details
Published in:Nucleic acids research 1982-04, Vol.10 (7), p.2177-2187
Main Authors: 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.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Cattle
Chymosin - genetics
Cloning, Molecular
DNA
DNA Restriction Enzymes
Enzyme Precursors - genetics
Protein Biosynthesis
RNA, Messenger - genetics
Stomach - enzymology
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Summary: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.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/10.7.2177