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PMID: 3539103 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Overproduction of the cyclic AMP receptor protein of Escherichia coli and expression of the engineered C-terminal DNA-binding domain.

The Biochemical journal ·Vol. 236 ·No. 3 ·1986-06-15 ·Pages 643-9

Gronenborn AM, Clore GM

Abstract

Overproduction of the cyclic AMP receptor protein (CRP) from Escherichia coli, up to 25% of the soluble cell protein, has been achieved in an inducible host-vector system under transcriptional control of the lambda promoter PL. This system is ideally suited for large scale production and purification of CRP. In addition, a structural gene for the DNA-binding domain of CRP has been constructed. To this end the nucleotide sequence coding for the C-terminus was fused to the sequence coding for the first 10 N-terminal amino acids and cloned into suitable vectors. Good expression was achieved using the lambda PL promoter. The gene product, beta CRP, is recognized by anti-CRP antibodies.

MeSH Terms
Base Sequence DNA, Bacterial/genetics Electrophoresis, Polyacrylamide Gel Escherichia coli/genetics Gene Expression Regulation Genetic Engineering Immunoelectrophoresis Plasmids Receptors, Cyclic AMP/biosynthesis,genetics Transformation, Genetic
Chemicals
DNA, Bacterial Receptors, Cyclic AMP
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Gronenborn A M
Clore G M
References (28)
28 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. Redesigning enzyme structure by site-directed mutagenesis: tyrosyl tRNA synthetase and ATP binding.
    Nature. 1982 Oct 21;299(5885):756-8 PMID: 6811955
  3. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  4. Construction of plasmid cloning vehicles that promote gene expression from the bacteriophage lambda pL promoter.
    Gene. 1979 Jan;5(1):59-76 PMID: 372049
  5. Structure of catabolite gene activator protein at 2.9 A resolution suggests binding to left-handed B-DNA.
    Nature. 1981 Apr 30;290(5809):744-9 PMID: 6261152
  6. Two helix DNA binding motif of CAP found in lac repressor and gal repressor.
    Nucleic Acids Res. 1982 Aug 25;10(16):5085-102 PMID: 6897114
  7. Proposed alpha-helical super-secondary structure associated with protein-dna recognition.
    J Mol Biol. 1982 Aug 25;159(4):745-51 PMID: 6897265
  8. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers.
    Gene. 1982 Oct;19(3):259-68 PMID: 6295879
  9. Autoregulation of the Escherichia coli crp gene: CRP is a transcriptional repressor for its own gene.
    Cell. 1983 Jan;32(1):141-9 PMID: 6297782
  10. Directed mutagenesis of dihydrofolate reductase.
    Science. 1983 Nov 18;222(4625):782-8 PMID: 6356360
  11. Deletion mutagenesis using an 'M13 splint': the N-terminal structural domain of tyrosyl-tRNA synthetase (B. stearothermophilus) catalyses the formation of tyrosyl adenylate.
    EMBO J. 1983;2(10):1827-9 PMID: 6315404
  12. A nuclear Overhauser enhancement study on the imino proton resonances of a DNA pentadecamer comprising the specific target site of the cyclic AMP receptor protein in the ara BAD operon.
    FEBS Lett. 1984 Jan 9;165(2):216-22 PMID: 6319182
  13. Vectors bearing a hybrid trp-lac promoter useful for regulated expression of cloned genes in Escherichia coli.
    Gene. 1983 Nov;25(2-3):167-78 PMID: 6363212
  14. A nuclear-Overhauser-enhancement study of the solution structure of a double-stranded DNA undecamer comprising a portion of the specific target site for the cyclic-AMP-receptor protein in the gal operon. Sequential resonance assignment.
    Eur J Biochem. 1984 May 15;141(1):119-29 PMID: 6327302
  15. Probing the catalytic mechanism of yeast triose phosphate isomerase by site-specific mutagenesis.
    Biochem Soc Trans. 1984 Apr;12(2):229-32 PMID: 6373437
  16. Interproton distance measurements in solution for a double-stranded DNA undecamer comprising a portion of the specific target site for the cycliC AMP receptor protein in the gal operon. A nuclear Overhauser enhancement study.
    FEBS Lett. 1984 Sep 17;175(1):117-23 PMID: 6479329
  17. Regulation of expression of the crp gene of Escherichia coli K-12: in vivo study.
    J Bacteriol. 1985 Jan;161(1):454-7 PMID: 2981809
  18. Altered Cro repressors from engineered mutagenesis of a synthetic cro gene.
    Proc Natl Acad Sci U S A. 1985 Feb;82(4):1084-8 PMID: 3156377
  19. The solution structure of a B-DNA undecamer comprising a portion of the specific target site for the cAMP receptor protein in the gal operon. Refinement on the basis of interproton distance data.
    EMBO J. 1985 Mar;4(3):829-35 PMID: 3891324
  20. Structure of the cro repressor from bacteriophage lambda and its interaction with DNA.
    Nature. 1981 Apr 30;290(5809):754-8 PMID: 6452580
  21. Plasmid vectors for high-efficiency expression controlled by the PL promoter of coliphage lambda.
    Gene. 1981 Oct;15(1):81-93 PMID: 6271633
  22. Molecular cloning and nucleotide sequencing of the gene for E. coli cAMP receptor protein.
    Nucleic Acids Res. 1982 Feb 25;10(4):1345-61 PMID: 6280140
  23. Cloning and sequence of the crp gene of Escherichia coli K 12.
    Nucleic Acids Res. 1982 Feb 25;10(4):1363-78 PMID: 6280141
  24. The operator-binding domain of lambda repressor: structure and DNA recognition.
    Nature. 1982 Jul 29;298(5873):443-7 PMID: 7088190
  25. Homology among DNA-binding proteins suggests use of a conserved super-secondary structure.
    Nature. 1982 Jul 29;298(5873):447-51 PMID: 6896364
  26. Structural similarity in the DNA-binding domains of catabolite gene activator and cro repressor proteins.
    Proc Natl Acad Sci U S A. 1982 May;79(10):3097-100 PMID: 6212926
  27. Structure of catabolite gene activator protein at 2.9-A resolution. Incorporation of amino acid sequence and interactions with cyclic AMP.
    J Biol Chem. 1982 Aug 25;257(16):9518-24 PMID: 6286624
  28. The purification and analysis of mechanism of action of a cyclic AMP-receptor protein from Escherichia coli.
    Methods Enzymol. 1974;38:367-76 PMID: 4375769
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2360643
Published
1986-06-15
Pages
643-9
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1146894
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2360643 PMC Full Text View journal details
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