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PLoS One ; 13(3): e0194420, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29543899

RESUMO

Interactions between DNA and proteins located in the cell nucleus play an important role in controlling physiological processes by specifying, augmenting and regulating context-specific transcription events. Chromatin immunoprecipitation (ChIP) is a widely used methodology to study DNA-protein interactions and has been successfully used in various cell types for over three decades. More recently, by combining ChIP with genomic screening technologies and Next Generation Sequencing (e.g. ChIP-seq), it has become possible to profile DNA-protein interactions (including covalent histone modifications) across entire genomes. However, the applicability of ChIP-chip and ChIP-seq has rarely been extended to non-model species because of a number of technical challenges. Here we report a method that can be used to identify genome wide covalent histone modifications in a group of non-model fruit fly species (Diptera: Tephritidae). The method was developed by testing and refining protocols that have been used in model organisms, including Drosophila melanogaster. We demonstrate that this method is suitable for a group of economically important pest fruit fly species, viz., Bactrocera dorsalis, Ceratitis capitata, Zeugodacus cucurbitae and Bactrocera tryoni. We also report an example ChIP-seq dataset for B. tryoni, providing evidence for histone modifications in the genome of a tephritid fruit fly for the first time. Since tephritids are major agricultural pests globally, this methodology will be a valuable resource to study taxa-specific evolutionary questions and to assist with pest management. It also provides a basis for researchers working with other non-model species to undertake genome wide DNA-protein interaction studies.


Assuntos
Imunoprecipitação da Cromatina/métodos , Drosophila melanogaster/metabolismo , Código das Histonas , Tephritidae/metabolismo , Animais , Ceratitis capitata/classificação , Ceratitis capitata/genética , Ceratitis capitata/metabolismo , DNA/genética , DNA/metabolismo , Dípteros/classificação , Dípteros/genética , Dípteros/metabolismo , Drosophila melanogaster/genética , Genoma de Inseto/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Histonas/metabolismo , Proteínas de Insetos/metabolismo , Ligação Proteica , Especificidade da Espécie , Tephritidae/classificação , Tephritidae/genética
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