Method for enrichment and isolation of endogenous transcription factor and complexes thereof and corresponding tandem arrays of concatenated transcription factor response elements
US8846884B2 · kind B2 · utility
Assignees
Inventors
Key dates
| Filing date | Dec 28, 2012 |
| Grant date | Sep 30, 2014 |
| Priority date | — |
| Expiry date | Jan 12, 2033 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC07K14/4702
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
The present invention provides a method for enrichment and isolation of endogenous transcription factors and their complexes. Also, this invention provides corresponding tandem arrays of concatenated transcription factor response elements (catTFRE). The method employs the property of transcription factors binding to sequence-specific DNA elements during regulation of gene expression. The catTFREs are designed and synthesized as concatenate dual copies of DNA response elements for various transcription factors. The DNA sequence of synthesized catTFRE is cloned to a target vector. Biotinylated catTFRE with 200 bp arms is prepared by PCR strategy. For enrichment and isolation of endogenous transcription factors and their complexes, the biotinylated catTFRE is immobilized to streptavidin-coated magnetic beads and then incubated with nuclear extract. Thereby endogenous transcription factors and their complexes are isolated from nuclear extract. Identification by mass spectrometry or other functional characterization can be further performed according to the application purposes.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.