Genetic Selection for Genes Encoding Sequence-Specific DNA-Binding Proteins

We describe a genetic selection method designed to facilitate the cloning of genes encoding sequence-specific DNA-binding proteins. The strategy selects for clones expressing particular sequence-specific DNA-binding activities from a library of clones encoding other, nonspecific proteins. Specific D...

Full description

Saved in:
Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1989-05, Vol.86 (10), p.3689-3693
Main Authors: Elledge, Stephen J., Sugiono, Paul, Guarente, Leonard, Davis, Ronald W.
Format: Article
Language:English
Subjects:
Animals
Binding sites
Biological and medical sciences
Biotechnology
Carrier proteins
Cloning, Molecular
DNA
DNA - genetics
DNA binding proteins
DNA-Binding Proteins - genetics
Escherichia coli - genetics
Fundamental and applied biological sciences. Psychology
Genes
Genetic engineering
Genetic screening
Genetic selection
Genetic technics
Libraries
Methods. Procedures. Technologies
Oligonucleotides
Plasmids
Promoter Regions, Genetic
Recombinant Fusion Proteins - metabolism
Recombinant Proteins - metabolism
Repressor Proteins - genetics
Selection, Genetic
Transcription Factors - genetics
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We describe a genetic selection method designed to facilitate the cloning of genes encoding sequence-specific DNA-binding proteins. The strategy selects for clones expressing particular sequence-specific DNA-binding activities from a library of clones encoding other, nonspecific proteins. Specific DNA-binding sites have been placed near the start of transcription of the strong synthetic conII promoter to create promoters that can be repressed by the corresponding sequence-specific DNA-binding proteins. Transcription from the conII derivatives in the absence of repression interferes with the phenotypic expression of an adjacent drug-resistance gene, aadA. Sequence-specific DNA-binding proteins are shown to repress these promoters and alleviate transcriptional interferance of aadA, resulting in drug resistance in cells expressing the appropriate DNA-binding protein.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.86.10.3689