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PMID: 3532112 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Promoters selected from random DNA sequences.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 19 ·1986-10-00 ·Pages 7405-9

Horwitz MS, Loeb LA

Abstract

We have selected a group of Escherichia coli promoters from random DNA sequences by replacing 19 base pairs at the -35 promoter region of the tetracycline resistance gene tetr of the plasmid pBR322. Substitution of 19 base pairs with chemically synthesized random sequences results in a maximum of 4(19) (about 3 X 10(11)) possible replacement sequences. From a population of about 1000 bacteria harboring plasmids with these random substitutions, tetracycline selection has revealed several functional -35 promoter sequences. These promoters have retained only partial homology to the -35 promoter consensus sequence. In three of these promoters, the consensus alignment shifts 10 nucleotides downstream, allowing the RNA polymerase to recognize another Pribnow box from within the original pBR322 sequence. Two of the sequences promote transcription more strongly than the native promoter. This technique may have application for the selection of additional DNA sequences with varied biological activity.

MeSH Terms

Base Sequence Chromosome Mapping DNA, Bacterial/genetics Escherichia coli/genetics Gene Expression Regulation Promoter Regions, Genetic Tetracycline/pharmacology Transcription, Genetic

Chemicals

DNA, Bacterial Tetracycline

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Horwitz M S

Loeb L A

References (27)

27 references, click to expand

Complete nucleotide sequence of the Escherichia coli plasmid pBR322.
Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:77-90 PMID: 383387
Eukaryotic DNA fragments which act as promoters for a plasmid gene.
Nature. 1979 Jan 25;277(5694):324-5 PMID: 368644
Construction and characterization of E. coli promoter-probe plasmid vectors. I. Cloning of promoter-containing DNA fragments.
Gene. 1979 Nov;7(3-4):271-88 PMID: 118083
Regulatory sequences involved in the promotion and termination of RNA transcription.
Annu Rev Genet. 1979;13:319-53 PMID: 94251
Deoxyribonucleic acid sequence of araBAD promoter mutants of Escherichia coli.
J Bacteriol. 1980 May;142(2):659-67 PMID: 6247327
Active efflux of tetracycline encoded by four genetically different tetracycline resistance determinants in Escherichia coli.
Proc Natl Acad Sci U S A. 1980 Jul;77(7):3974-7 PMID: 7001450
lac Promoter mutation Pr115 generates a new transcription initiation point.
J Mol Biol. 1980 May 25;139(3):551-6 PMID: 6160250
Organization of transcriptional signals in plasmids pBR322 and pACYC184.
Proc Natl Acad Sci U S A. 1981 Jan;78(1):167-71 PMID: 7017708
Synthesis of oligoguanylates on oligocytidylate templates.
J Mol Evol. 1981;17(5):295-302 PMID: 7277512
Automated synthesis of gene fragments.
Science. 1981 Oct 16;214(4518):270-4 PMID: 6169150
A set of synthetic oligodeoxyribonucleotide primers for DNA sequencing in the plasmid vector pBR322.
Gene. 1981 Dec;16(1-3):21-6 PMID: 6282692
Precise location of two promoters for the beta-lactamase gene of pBR322. S1 mapping of ribonucleic acid isolated from Escherichia coli or synthesized in vitro.
J Biol Chem. 1982 Aug 10;257(15):9205-10 PMID: 6178738
Construction and characterization of E. coli promoter-probe plasmid vectors. III. pBR322 derivatives with deletions in the tetracycline resistance promoter region.
Gene. 1982 Dec;20(2):291-304 PMID: 6299894
Targeted random mutagenesis: the use of ambiguously synthesized oligonucleotides to mutagenize sequences immediately 5' of an ATG initiation codon.
Nucleic Acids Res. 1983 May 25;11(10):3113-21 PMID: 6304623
Compilation and analysis of Escherichia coli promoter DNA sequences.
Nucleic Acids Res. 1983 Apr 25;11(8):2237-55 PMID: 6344016
Studies on transformation of Escherichia coli with plasmids.
J Mol Biol. 1983 Jun 5;166(4):557-80 PMID: 6345791
Tetracycline resistance determined by pBR322 is mediated by one polypeptide.
Gene. 1983 Dec;26(2-3):197-203 PMID: 6323260
Plasmid vectors for the selection of promoters.
Gene. 1984 Feb;27(2):151-60 PMID: 6327464
Mechanism and control of transcription initiation in prokaryotes.
Annu Rev Biochem. 1985;54:171-204 PMID: 3896120
Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes.
Cell. 1986 Jan 31;44(2):283-92 PMID: 3943125
Nucleotide sequence of an RNA polymerase binding site from the DNA of bacteriophage fd.
Proc Natl Acad Sci U S A. 1975 Feb;72(2):737-41 PMID: 1054851
Bacteriophage T7 early promoters: nucleotide sequences of two RNA polymerase binding sites.
J Mol Biol. 1975 Dec 15;99(3):419-43 PMID: 765476
Sequence of promoter for coat protein gene of bacteriophage fd.
Nature. 1976 Mar 25;260(5549):297-302 PMID: 1256568
Interaction of RNA polymerase with promoters from bacteriophage fd.
Eur J Biochem. 1977 Mar 15;74(1):107-13 PMID: 300680
DNA sequencing with chain-terminating inhibitors.
Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
The expression of tetracycline resistance after insertion of foreign DNA fragments between the EcoRI and HindIII sites of the plasmid cloning vector pBR 322.
Mol Gen Genet. 1978 Jul 25;163(3):301-5 PMID: 355856
Characterizing wild-type and mutant promoters of the tetracycline resistance gene in pBR313.
Nucleic Acids Res. 1979 Jul 25;6(10):3267-87 PMID: 384366

Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.83.19.7405

Published

1986-10-00

Pages

7405-9

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC386726

Subset

External Links

PubMed source DOI: 10.1073/pnas.83.19.7405 PMC Full Text View journal details

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