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  • Methylome of DNase I sensitive chromatin in Populus trichocarpa shoot apical meristematic cells: a simplified approach revealing characteristics of gene-body DNA methylation in open chromatin state.

Methylome of DNase I sensitive chromatin in Populus trichocarpa shoot apical meristematic cells: a simplified approach revealing characteristics of gene-body DNA methylation in open chromatin state.

The New phytologist (2012-12-21)
Clément Lafon-Placette, Patricia Faivre-Rampant, Alain Delaunay, Nathaniel Street, Franck Brignolas, Stéphane Maury
ABSTRACT

DNA methylation is involved in the control of plant development and adaptation to the environment through modifications of chromatin compaction and gene expression. In poplar (Populus trichocarpa), a perennial plant, variations in DNA methylation have been reported between genotypes and tissues or in response to drought. Nevertheless, the relationships between gene-body DNA methylation, gene expression and chromatin compaction still need clarification. Here, DNA methylation was mapped in the noncondensed chromatin fraction from P. trichocarpa shoot apical meristematic cells, the center of plant morphogenesis, where DNA methylation variations could influence the developmental trajectory. DNase I was used to isolate the noncondensed chromatin fraction. Methylated sequences were immunoprecipitated, sequenced using Illumina/Solexa technology and mapped on the v2.0 poplar genome. Bisulfite sequencing of candidate sequences was used to confirm mapping data and to assess cytosine contexts and methylation levels. While the methylated DNase I hypersensitive site fraction covered 1.9% of the poplar genome, it contained sequences corresponding to 74% of poplar gene models, mostly exons. The level and cytosine context of gene-body DNA methylation varied with the structural characteristics of the genes. Taken together, our data show that DNA methylation is widespread and variable among genes in open chromatin of meristematic cells, in agreement with a role in their developmental trajectory.

MATERIALS
Product Number
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Product Description

Sigma-Aldrich
Deoxyribonuclease I RNase-free solution from bovine pancreas
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Deoxyribonuclease I bovine, recombinant, expressed in Pichia pastoris, buffered aqueous glycerol solution, ≥5,000 units/mg protein
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Deoxyribonuclease I from bovine pancreas, Standardized vial containing 2,000 Kunitz units of DNase I (D4527), vial of ≥0.25 mg total protein
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Deoxyribonuclease I from bovine pancreas, Type II, lyophilized powder, Protein ≥80 %, ≥2,000 units/mg protein
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Deoxyribonuclease I from bovine pancreas, lyophilized powder, Protein ≥85 %, ≥400 Kunitz units/mg protein
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Deoxyribonuclease I from bovine pancreas, Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein
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Deoxyribonuclease I from bovine pancreas, Type II-S, lyophilized powder, Protein ≥80 %, ≥2,000 units/mg protein
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Deoxyribonuclease I bovine, recombinant, expressed in Pichia pastoris, lyophilized powder, RNAse and protease, free