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Methods Mol Biol ; 2819: 39-53, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-39028501

RESUMEN

Nucleotide sequences recognized and bound by DNA-binding proteins (DBPs) are critical to controlling and maintaining gene expression, replication, chromosome segregation, cell division, and nucleoid structure in bacterial cells. Therefore, determination of the binding sequences of DBPs is important not only to study DBP recognition mechanisms but also to understand the fundamentals of cell homeostasis. While ChIP-seq analysis appears to be an effective way to determine DBP binding sites on the genome, the resolution is sometimes not sufficient to identify the sites precisely. Here we introduce a simple and effective method named Genome Footprinting with high-throughput sequencing (GeF-seq) to determine binding sites of DBPs with single base-pair resolution. GeF-seq detects binding sites of DBPs as sharp peaks and thus makes it possible to identify the recognition sequence in each "binding peak" more easily and accurately compared to the common ChIP-seq.


Asunto(s)
Secuenciación de Inmunoprecipitación de Cromatina , Proteínas de Unión al ADN , Secuenciación de Nucleótidos de Alto Rendimiento , Secuenciación de Inmunoprecipitación de Cromatina/métodos , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Sitios de Unión , Proteínas de Unión al ADN/metabolismo , Proteínas de Unión al ADN/genética , Emparejamiento Base , Unión Proteica , Huella de ADN/métodos
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