Abstract
Protein-DNA interactions are important in determining the transcriptional status of chromatin and in determining the tissue-specificity, timing and quantitative levels of transcription. Substantial effort is being directed towards understanding the mechanisms of gene expression in plants, and isolating the components involved, such as transcription factors. Avariety of methods have been applied to the study of interactions between putative transcription factors and gene promoters in plants, but approaches that permit examination of protein-DNA interactions in intact plant tissues offer a more direct examination of these interactions in differentiated plant tissues. Here we describe in detail the method of ligation-mediated PCR forin-vivo footprinting, which can be used to study protein-DNA interactionsin planta with a high degree of precision and sensitivity.
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Abbreviations
- OP-Cu:
-
1,10-o-phenanthroline cuprous complex
- DMS:
-
dimethylsulphate
- LMPCR:
-
ligation-mediated PCR
- LMWG-1D1:
-
gene encoding low-molecular-weight glutenin
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Hammond-Kosack, M.C.U., Bevan, M.W. A practical guide to ligation-mediated PCR footprinting andin-vivo DNA analysis using plant tissues. Plant Mol Biol Rep 11, 249–272 (1993). https://doi.org/10.1007/BF02669852
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DOI: https://doi.org/10.1007/BF02669852