Identifying DNA sequences recognized by a transcription factor using a bacterial one-hybrid system.

Identifying DNA sequences recognized by a transcription factor using a bacterial one-hybrid system. Research Abstract Details 

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Identifying DNA sequences recognized by a transcription factor using a bacterial one-hybrid system. Abstract Text:

Xiangdong Meng,Scot A Wolfe,Xiangdong Meng,Scot A Wolfe,

Bacterial-based interaction trap systems provide a powerful method to identify interacting macromolecules. When carried out in the context of a genetic selection, interacting pairs can be rapidly isolated from large combinatorial libraries. This technology has been adapted to allow the identification of DNA-binding sequences for a transcription factor (TF) from a large randomized library. This procedure uses a library of randomized binding sites upstream of a cocistronic HIS3-URA3 reporter cassette. The URA3 reporter allows self-activating sequences to be removed from the library through counter-selection. The HIS3 reporter allows sequences that are recognized by a TF to be isolated from the library, where transcriptional activation is mediated by fusion of the TF to the alpha-subunit of RNA polymerase. This technology can be used to characterize monomeric, homodimeric and heterodimeric DNA-binding domains and, once a suitable library is constructed, binding sites can be identified in approximately 10 d. The bacterial one-hybrid system allows larger libraries to be searched than the corresponding yeast one-hybrid system and, unlike SELEX, it does not require purification of the TF(s). The complexity of the binding site libraries that can be searched using the bacterial system is, however, more limited than SELEX, and some eukaryotic factors may not express or fold efficiently in the bacterial system.

Identifying DNA sequences recognized by a transcription factor using a bacterial one-hybrid system. Publishing Authors By Initials

X Meng,SA Wolfe,X Meng,SA Wolfe,

For similar investigative techniques: genetic techniques: cloning, molecular: two-hybrid system techniques research abstracts see: investigative techniques: genetic techniques: cloning, molecular: two-hybrid system techniques research

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Identifying DNA sequences recognized by a transcription factor using a bacterial one-hybrid system. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Nature protocols

VOLUME: 1

Page Numbers: 30-45

Journal Abbreviation:

ISSN: 1750-2799

DAY: 3

MONTH: 12

YEAR: 2006

Identifying DNA sequences recognized by a transcription factor using a bacterial one-hybrid system. Information

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LANGUAGE: eng

NlmUniqueID: 101284307

Identifying DNA sequences recognized by a transcription factor using a bacterial one-hybrid system. Keywords Mesh Terms:

KEYWORDS: Two-Hybrid System Techniques

MESH TERMS: metabolism

Chemical & Substance for Abstract: Identifying DNA sequences recognized by a transcription factor using a bacterial one-hybrid system. Information

Substance Name: Transcription Factors

Registry Number: 0

Grant and Affiliation Information for Identifying DNA sequences recognized by a transcription factor using a bacterial one-hybrid system.

AFFILIATION: Program in Gene Function and Expression, University of Massachusetts Medical School, 364 Plantation Street, Worcester, Massachusetts 01605, USA.

Country: England

AGENCY: United States NIGMS

GRANT: 1R01GM068110

ACRONYM: GM

MEDLINETA: Nat Protoc

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