Method for Cloning In Vivo Targets of the Egr-1 Transcription Factor | Zendy

Ian de Belle | Zendy; Dan Mercola | Zendy; Eileen D. Adamson | Zendy

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Method for Cloning In Vivo Targets of the Egr-1 Transcription Factor

Author(s) -

Ian de Belle,

Dan Mercola,

Eileen D. Adamson

Publication year - 2000

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/00291rr03

Subject(s) - biology , gene , transcription factor , cloning (programming) , transcription (linguistics) , dna , in vivo , promoter , microbiology and biotechnology , genetics , gene expression , computational biology , linguistics , philosophy , computer science , programming language

A methodology is described that allows the in vivo trapping of transcription factors to their target regulatory elements in multiple genes simultaneously. Cross-linking using formaldehyde is the first of several steps to isolate, purify, clone and characterize multiple gene promoter DNA fragments. The example that we use indicates that the TGFβ1 gene is a direct target induced by Egr-1 in HT1080 cells that express constitutive Egr-1, thus explaining the growth retardation that follows Egr-1 expression. The genes identified using this procedure reflect the specific activities of Egr-1 at that moment in the cell and provide a method for confirmation of genes that are the direct targets of Egr-1 action.

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