RESUMO
Nucleosome occupancy plays an important role in chromatin compaction, affecting biological processes by hampering the binding of cis-acting elements such as transcription factors, RNA polymerase machinery, and coregulatory. Accessible regions allow for cis-acting elements to bind DNA and regulate transcription. Here, we detail our protocol to profile nucleosome occupancy and chromatin structure dynamics under drought stress at the genome-wide scale using micrococcal nuclease (MNase) digestion. Combining variable MNase concentration treatments and high-throughput sequencing, we investigate the changes in the overall chromatin state using bread wheat samples from an exemplary drought experiment.
Assuntos
Sequenciamento de Nucleotídeos em Larga Escala , Nuclease do Micrococo , Nucleossomos , Estresse Fisiológico , Triticum , Nucleossomos/metabolismo , Nucleossomos/genética , Nuclease do Micrococo/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Estresse Fisiológico/genética , Triticum/genética , Triticum/metabolismo , Secas , Regulação da Expressão Gênica de Plantas , Cromatina/metabolismo , Cromatina/genética , Plantas/genética , Plantas/metabolismoRESUMO
We have developed a tool for controlled expression of heterologous or ectopic genes in the chestnut pathogen Cryphonectria parasitica using the promoter region from a putative copper-regulated transporter gene. In addition, we have found that expression control via this system is not affected by the virulence-attenuating hypovirus CHV1-EP713.