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. 1985 Dec;164(3):1353–1355. doi: 10.1128/jb.164.3.1353-1355.1985

lac Up-promoter mutants with increased homology to the consensus promoter sequence.

W Mandecki, R A Goldman, B S Powell, M H Caruthers
PMCID: PMC219337  PMID: 2999082

Abstract

Four lac promoter mutants were constructed. The mutations increased the homology between the lac promoter and the consensus promoter sequences by introducing the consensus -10 and -35 regions and the consensus spacing of 17 residues between these two regions. The promoter mutants were cloned into a pBR322-derivatized vector upstream from the lacZ gene, and levels of beta-galactosidase were an indication of promoter activity. All mutants exhibited higher activity than did the wild-type promoter.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ackerson J. W., Gralla J. D. In vivo expression of lac promoter variants with altered -10, -35, and spacer sequences. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 1):473–476. doi: 10.1101/sqb.1983.047.01.056. [DOI] [PubMed] [Google Scholar]
  2. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Hopkins J. D. A new class of promoter mutations in the lactose operon of Escherichia coli. J Mol Biol. 1974 Aug 25;87(4):715–724. doi: 10.1016/0022-2836(74)90080-1. [DOI] [PubMed] [Google Scholar]
  4. Inouye S., Inouye M. Up-promoter mutations in the lpp gene of Escherichia coli. Nucleic Acids Res. 1985 May 10;13(9):3101–3110. doi: 10.1093/nar/13.9.3101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Mandecki W., Caruthers M. H. Mutants of the lac promoter with large insertions and deletions between the CAP binding site and the -35 region. Gene. 1984 Nov;31(1-3):263–267. doi: 10.1016/0378-1119(84)90219-1. [DOI] [PubMed] [Google Scholar]
  6. Mandecki W., Mollison K. W., Bolling T. J., Powell B. S., Carter G. W., Fox J. L. Chemical synthesis of a gene encoding the human complement fragment C5a and its expression in Escherichia coli. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3543–3547. doi: 10.1073/pnas.82.11.3543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Mandecki W., Reznikoff W. S. A lac promoter with a changed distance between -10 and -35 regions. Nucleic Acids Res. 1982 Feb 11;10(3):903–912. doi: 10.1093/nar/10.3.903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  9. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  10. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  11. Shapira S. K., Chou J., Richaud F. V., Casadaban M. J. New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequences encoding an enzymatically active carboxy-terminal portion of beta-galactosidase. Gene. 1983 Nov;25(1):71–82. doi: 10.1016/0378-1119(83)90169-5. [DOI] [PubMed] [Google Scholar]
  12. Siebenlist U., Simpson R. B., Gilbert W. E. coli RNA polymerase interacts homologously with two different promoters. Cell. 1980 Jun;20(2):269–281. doi: 10.1016/0092-8674(80)90613-3. [DOI] [PubMed] [Google Scholar]
  13. Silverstone A. E., Arditti R. R., Magasanik B. Catabolite-insensitive revertants of lac promoter mutants. Proc Natl Acad Sci U S A. 1970 Jul;66(3):773–779. doi: 10.1073/pnas.66.3.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Stefano J. E., Gralla J. D. Spacer mutations in the lac ps promoter. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1069–1072. doi: 10.1073/pnas.79.4.1069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. de Boer H. A., Comstock L. J., Vasser M. The tac promoter: a functional hybrid derived from the trp and lac promoters. Proc Natl Acad Sci U S A. 1983 Jan;80(1):21–25. doi: 10.1073/pnas.80.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. deHaseth P. L., Goldman R. A., Cech C. L., Caruthers M. H. Chemical synthesis and biochemical reactivity of bacteriophage lambda PR promoter. Nucleic Acids Res. 1983 Feb 11;11(3):773–787. doi: 10.1093/nar/11.3.773. [DOI] [PMC free article] [PubMed] [Google Scholar]

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