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A simplified method for the analysis of transcription factor-promoter interactions that allows high-throughput data generation.
- Source :
-
The Plant journal : for cell and molecular biology [Plant J] 2007 Jun; Vol. 50 (5), pp. 911-6. Date of Electronic Publication: 2007 Apr 08. - Publication Year :
- 2007
-
Abstract
- Transient expression systems are intensively used to study the transactivation potential of transcription factors and to confirm target promoters. Here we present a novel system based on the high-efficiency transformation of cultured Arabidopsis thaliana cells by agrobacteria. To demonstrate the potential of this system, we compared it with a commonly used protoplast transfection assay, and studied the regulation of phenylpropanoid biosynthetic pathway genes by various transcription factors. Both systems led to comparable results on the regulation of the promoters tested. However, the agrobacterium-mediated co-transformation assay needs significantly less time, requires only mixing of cultured plant cells with agrobacteria, is less labour-intensive and allows handling of multiple assays in parallel, making it suitable for medium- to high-throughput analyses. In addition, the binary vectors used are the same for both cell-based assays and stable plant transformations.
- Subjects :
- Arabidopsis genetics
Arabidopsis metabolism
Arabidopsis microbiology
Arabidopsis Proteins genetics
Arabidopsis Proteins metabolism
Bacterial Proteins genetics
Cloning, Molecular methods
Glucuronidase genetics
Protoplasts
Rhizobium enzymology
Rhizobium genetics
Transfection methods
Promoter Regions, Genetic
Transcription Factors metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0960-7412
- Volume :
- 50
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- The Plant journal : for cell and molecular biology
- Publication Type :
- Academic Journal
- Accession number :
- 17425717
- Full Text :
- https://doi.org/10.1111/j.1365-313X.2007.03088.x