| Chapter title |
Rapid and Quantitative CELD Assay to Measure the Specificity of Transcription Factor-DNA-Binding Interactions and Identify cis-Elements
|
|---|---|
| Chapter number | 18 |
| Book title |
Plant Transcription Factors
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-8657-6_18 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-8656-9, 978-1-4939-8657-6
|
| Authors |
Sundaravelpandian Kalaipandian, Gang-Ping Xue, Kalaipandian, Sundaravelpandian, Xue, Gang-Ping |
| Abstract |
Specific interaction between transcription factor (TF) and cis-regulatory elements in the context of chromatin is the key to determining gene expression. Thus, it is important to measure DNA-binding specificity and identify cis-elements of TFs of your interest. In this chapter, we described a microwell-based assay to determine DNA-binding specificity by using translational fusion of a TF with a highly active cellulase (CELD), which hydrolyzes 4-methylumbelliferyl β-D-cellobioside to a fluorescent 4-methylumbelliferone product. The hydrolysis activity arrows us to quantify the binding strength between TF-CELD fusion protein and biotinylated DNA sequences in a 96-well microplate. The high-throughput and quantitative nature of CELD assay enable researchers to test a large number of putative DNA-binding sites of TFs, which subsequently leads to the identification of its direct target genes. |
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Mendeley readers
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| Unknown | 2 | 100% |
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|---|---|---|
| Researcher | 1 | 50% |
| Student > Master | 1 | 50% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 1 | 50% |
| Agricultural and Biological Sciences | 1 | 50% |