Method for Cloning In Vivo Targets of the Egr-1 Transcription Factor
| Autor: | I de Belle, Eileen D. Adamson, D Mercola |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Transcriptional Activation
Transcription Genetic Fibrosarcoma Response element Gene Expression Breast Neoplasms Molecular cloning Biology Transfection Polymerase Chain Reaction Sensitivity and Specificity General Biochemistry Genetics and Molecular Biology Immediate early protein Immediate-Early Proteins Transforming Growth Factor beta Transcription (biology) Formaldehyde Tumor Cells Cultured Humans Cloning Molecular Gene Transcription factor Immunosorbent Techniques Early Growth Response Protein 1 Cloning Binding Sites Promoter DNA Molecular biology Cell biology DNA-Binding Proteins body regions Cross-Linking Reagents Tetradecanoylphorbol Acetate hormones hormone substitutes and hormone antagonists Transcription Factors Biotechnology |
| Zdroj: | BioTechniques. 29:162-169 |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/00291rr03 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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