Search tips
Search criteria 


Logo of narLink to Publisher's site
Nucleic Acids Res. 1982 April 10; 10(7): 2177–2187.
PMCID: PMC320601

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


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.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (937K), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Foltmann B, Pedersen VB, Kauffman D, Wybrandt G. The primary structure of calf chymosin. J Biol Chem. 1979 Sep 10;254(17):8447–8456. [PubMed]
  • Foltmann B, Pedersen VB, Jacobsen H, Kauffman D, Wybrandt G. The complete amino acid sequence of prochymosin. Proc Natl Acad Sci U S A. 1977 Jun;74(6):2321–2324. [PubMed]
  • Davis BD, Tai PC. The mechanism of protein secretion across membranes. Nature. 1980 Jan 31;283(5746):433–438. [PubMed]
  • Pedersen VB, Christensen KA, Foltmann B. Investigations on the activation of bovine prochymosin. Eur J Biochem. 1979 Mar;94(2):573–580. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Bonner WM, Laskey RA. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. [PubMed]
  • Harris TJ, Brown F, Sangar DV. Differential precipitation of foot and mouth disease virus proteins made in vivo and in vitro by hyperimmune and virus particle guinea pig antisera. Virology. 1981 Jul 15;112(1):91–98. [PubMed]
  • Retzel EF, Collett MS, Faras AJ. Enzymatic synthesis of deoxyribonucleic acid by the avian retrovirus reverse transcriptase in vitro: optimum conditions required for transcription of large ribonucleic acid templates. Biochemistry. 1980 Feb 5;19(3):513–518. [PubMed]
  • Hoeijmakers JH, Borst P, van den Burg J, Weissmann C, Cross GA. The isolation of plasmids containing DNA complementary to messenger RNA for variant surface glycoproteins of Trypanosoma brucei. Gene. 1980 Mar;8(4):391–417. [PubMed]
  • Otsuka A. Recovery of DNA fragments inserted by the "tailing" method: regeneration of PstI restriction sites. Gene. 1981 May;13(4):339–346. [PubMed]
  • Morrison DA. Transformation and preservation of competent bacterial cells by freezing. Methods Enzymol. 1979;68:326–331. [PubMed]
  • Gergen JP, Stern RH, Wensink PC. Filter replicas and permanent collections of recombinant DNA plasmids. Nucleic Acids Res. 1979 Dec 20;7(8):2115–2136. [PMC free article] [PubMed]
  • Hanahan D, Meselson M. Plasmid screening at high colony density. Gene. 1980 Jun;10(1):63–67. [PubMed]
  • Edge MD, Green AR, Heathcliffe GR, Meacock PA, Schuch W, Scanlon DB, Atkinson TC, Newton CR, Markham AF. Total synthesis of a human leukocyte interferon gene. Nature. 1981 Aug 20;292(5825):756–762. [PubMed]
  • Land H, Grez M, Hauser H, Lindenmaier W, Schütz G. 5'-Terminal sequences of eucaryotic mRNA can be cloned with high efficiency. Nucleic Acids Res. 1981 May 25;9(10):2251–2266. [PMC free article] [PubMed]
  • Ish-Horowicz D, Burke JF. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. [PMC free article] [PubMed]
  • Maxam AM, Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. [PubMed]
  • Uchiyama H, Uozumi T, Beppu T, Arima K. Purification of prorennin and production of its antibody. J Biochem. 1981 Aug;90(2):483–487. [PubMed]
  • Nishimori K, Kawaguchi Y, Hidaka M, Uozumi T, Beppu T. Cloning in Escherichia coli of the structural gene of prorennin, the precursor of calf milk-clotting enzyme renin. J Biochem. 1981 Sep;90(3):901–904. [PubMed]
  • Boothroyd JC, Highfield PE, Cross GA, Rowlands DJ, Lowe PA, Brown F, Harris TJ. Molecular cloning of foot and mouth disease virus genome and nucleotide sequences in the structural protein genes. Nature. 1981 Apr 30;290(5809):800–802. [PubMed]
  • Quinto C, Quiroga M, Swain WF, Nikovits WC, Jr, Standring DN, Pictet RL, Valenzuela P, Rutter WJ. Rat preprocarboxypeptidase A: cDNA sequence and preliminary characterization of the gene. Proc Natl Acad Sci U S A. 1982 Jan;79(1):31–35. [PubMed]
  • Kozak M. Possible role of flanking nucleotides in recognition of the AUG initiator codon by eukaryotic ribosomes. Nucleic Acids Res. 1981 Oct 24;9(20):5233–5252. [PMC free article] [PubMed]
  • Wain-Hobson S, Nussinov R, Brown RJ, Sussman JL. Preferential codon usage in genes. Gene. 1981 May;13(4):355–364. [PubMed]
  • Foltmann B, Pedersen VB. Comparison of the primary structures of acidic proteinases and of their zymogens. Adv Exp Med Biol. 1977;95:3–22. [PubMed]
  • Proudfoot NJ, Brownlee GG. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. [PubMed]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press