Home LiteratureArticle Details
PMID: 6184675 Published · ppublish English Journal Article

The construction of a synthetic Escherichia coli trp promoter and its use in the expression of a synthetic interferon gene.

Nucleic acids research ·Vol. 10 ·No. 21 ·1982-11-11 ·Pages 6639-57

Windass JD, Newton CR, De Maeyer-Guignard J, Moore VE, Markham AF, Edge MD

Abstract

An 82 base pair DNA fragment has been synthesised which contains the E. coli trp promoter and operator sequences and also encodes the first Shine Dalgarno sequence of the trp operon. This DNA fragment is flanked by EcoRI and ClaI/TaqI cohesive ends and is thus easy to clone, transfer between vector systems and couple to genes to drive their expression. It has been cloned into plasmid pAT153, producing a convenient trp promoter vector. We have also joined the fragment to a synthetic IFN-alpha 1 gene, using synthetic oligonucleotides to generate a completely natural, highly efficient bacterial translation initiation signal on the promoter proximal side of the IFN gene. Plasmids carrying this construction enable E. coli cells to express IFN-alpha 1 almost constitutively and with significantly higher efficiency than from a lacUV5 promoter based system.

MeSH Terms
Base Composition Base Sequence Cloning, Molecular DNA Restriction Enzymes Escherichia coli/genetics Genes, Synthetic Interferons/genetics Mutation Oligodeoxyribonucleotides/chemical synthesis,genetics Operon Plasmids Polynucleotide 5'-Hydroxyl-Kinase/metabolism T-Phages/enzymology
Chemicals
Oligodeoxyribonucleotides Interferons Polynucleotide 5'-Hydroxyl-Kinase DNA Restriction Enzymes
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Windass J D
Newton C R
De Maeyer-Guignard J
Moore V E
Markham A F
Edge M D
References (39)
39 references, click to expand
  1. Functional expression of cloned yeast DNA in Escherichia coli: specific complementation of argininosuccinate lyase (argH) mutations.
    J Mol Biol. 1978 Apr 25;120(4):517-32 PMID: 349166
  2. Total synthesis of a human leukocyte interferon gene.
    Nature. 1981 Aug 20;292(5825):756-62 PMID: 6167861
  3. Aberrant immunity behaviour of hybrid lambda imm21 phages containing the DNA of ColE1-type plasmids.
    Mol Gen Genet. 1979;172(3):329-37 PMID: 45615
  4. Recovery of DNA from gels.
    Methods Enzymol. 1980;65(1):371-80 PMID: 7374460
  5. The structure of eight distinct cloned human leukocyte interferon cDNAs.
    Nature. 1981 Mar 5;290(5801):20-6 PMID: 6163083
  6. Inhibition of lacZ gene translation initiation in trp-lac fusion strains.
    J Bacteriol. 1974 Mar;117(3):1231-9 PMID: 4591949
  7. Direct expression in Escherichia coli of a DNA sequence coding for human growth hormone.
    Nature. 1979 Oct 18;281(5732):544-8 PMID: 386136
  8. Human leukocyte interferon produced by E. coli is biologically active.
    Nature. 1980 Oct 2;287(5781):411-6 PMID: 6159538
  9. Thr region between the operator and first structural gene of the tryptophan operon of Escherichia coli may have a regulatory function.
    J Mol Biol. 1973 May 5;76(1):89-101 PMID: 4578102
  10. Synthesis of a model promoter for gene expression in Escherichia coli.
    Nucleic Acids Symp Ser. 1980;(7):365-76 PMID: 6265875
  11. Solid phase phosphotriester synthesis of large oligodeoxyribonucleotides on a polyamide support.
    Nucleic Acids Res. 1980 Nov 25;8(22):5193-205 PMID: 7465412
  12. Synthesis of human fibroblast interferon by E. coli.
    Nucleic Acids Res. 1980 Sep 25;8(18):4057-74 PMID: 6159584
  13. Human growth hormone: complementary DNA cloning and expression in bacteria.
    Science. 1979 Aug 10;205(4406):602-7 PMID: 377496
  14. Expression in Escherichia coli of a chemically synthesized gene for the hormone somatostatin.
    Science. 1977 Dec 9;198(4321):1056-63 PMID: 412251
  15. Antiviral activity of bacteria-derived human alpha interferons against encephalomyocarditis virus infection of mice.
    Infect Immun. 1982 Feb;35(2):660-5 PMID: 6173327
  16. Influenza antigenic determinants are expressed from haemagglutinin genes cloned in Escherichia coli.
    Nature. 1980 Jan 10;283(5743):171-4 PMID: 6153123
  17. A rapid boiling method for the preparation of bacterial plasmids.
    Anal Biochem. 1981 Jun;114(1):193-7 PMID: 6269464
  18. The complete nucleotide sequence of the tryptophan operon of Escherichia coli.
    Nucleic Acids Res. 1981 Dec 21;9(24):6647-68 PMID: 7038627
  19. Nucleotide sequence and expression of Escherichia coli trpR, the structural gene for the trp aporepressor.
    Proc Natl Acad Sci U S A. 1980 Dec;77(12):7117-21 PMID: 7012834
  20. Mutagenesis at a specific position in a DNA sequence.
    J Biol Chem. 1978 Sep 25;253(18):6551-60 PMID: 681366
  21. A bacterial clone synthesizing proinsulin.
    Proc Natl Acad Sci U S A. 1978 Aug;75(8):3727-31 PMID: 358198
  22. Rapid synthesis of oligodeoxyribonucleotides. IV. Improved solid phase synthesis of oligodeoxyribonucleotides through phosphotriester intermediates.
    Nucleic Acids Res. 1980 Mar 11;8(5):1081-96 PMID: 7443540
  23. Expression of the human fibroblast interferon gene in Escherichia coli.
    Proc Natl Acad Sci U S A. 1980 Sep;77(9):5230-3 PMID: 6159642
  24. Maximizing gene expression on a plasmid using recombination in vitro.
    Cell. 1978 Jan;13(1):65-71 PMID: 340049
  25. A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
    Nucleic Acids Res. 1979 Nov 24;7(6):1513-23 PMID: 388356
  26. Normal repression in a deletion mutant lacking almost the entire operator-proximal gene of the tryptophan operon of E. coli.
    Nat New Biol. 1972 May 10;237(71):47-9 PMID: 4555683
  27. Quantitative aspects of gene expression in a lambda-trp fusion operon.
    Mol Gen Genet. 1974 Apr 9;130(1):9-20 PMID: 4837659
  28. Directory of restriction endonucleases.
    Methods Enzymol. 1980;65(1):1-15 PMID: 6246329
  29. TRANSFORMATION OF BIOCHEMICALLY DEFICIENT STRAINS OF BACILLUS SUBTILIS BY DEOXYRIBONUCLEATE.
    Proc Natl Acad Sci U S A. 1958 Oct 15;44(10):1072-8 PMID: 16590310
  30. Target cell specificity of two species of human interferon-alpha produced in Escherichia coli and of hybrid molecules derived from them.
    Proc Natl Acad Sci U S A. 1981 May;78(5):2848-52 PMID: 6166941
  31. Synthesis of an ovalbumin-like protein by Escherichia coli K12 harbouring a recombinant plasmid.
    Nature. 1978 Oct 12;275(5680):505-10 PMID: 80751
  32. Trans-complementable copy-number mutants of plasmid ColE1.
    Nature. 1980 Jan 10;283(5743):216-8 PMID: 7350544
  33. Synthesis of simian virus 40 t antigen in Escherichia coli.
    Proc Natl Acad Sci U S A. 1979 Nov;76(11):5596-600 PMID: 230483
  34. A functional hybrid ribosome binding site in tryptophan operon messenger RNA of Escherichia coli.
    J Mol Biol. 1980 Nov 5;143(3):335-41 PMID: 6163864
  35. Properties of a human alpha-interferon purified from E. coli extracts.
    J Interferon Res. 1981;1(3):381-90 PMID: 6180070
  36. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
  37. Determinant of cistron specificity in bacterial ribosomes.
    Nature. 1975 Mar 6;254(5495):34-8 PMID: 803646
  38. Synthesis of hepatitis B surface and core antigens in E. coli.
    Nature. 1981 Jun 11;291(5815):503-6 PMID: 6262658
  39. Gene structure in the tryptophan operon of Escherichia coli. Nucleotide sequence of trpC and the flanking intercistronic regions.
    J Mol Biol. 1980 Oct 5;142(4):519-30 PMID: 7007653
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/10.21.6639
Published
1982-11-11
Pages
6639-57
Language
English
Region
England
NLM ID
0411011
PMCID
PMC326954
Subset
IM
Databases
GENBANK
J01714, M12471, M12472
External Links
PubMed source DOI: 10.1093/nar/10.21.6639 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2