Your browser doesn't support javascript.
loading
In vivo insertion pool sequencing identifies virulence factors in a complex fungal-host interaction.
Uhse, Simon; Pflug, Florian G; Stirnberg, Alexandra; Ehrlinger, Klaus; von Haeseler, Arndt; Djamei, Armin.
Afiliación
  • Uhse S; Gregor Mendel Institute (GMI), Austrian Academy of Sciences, Vienna BioCenter (VBC), Vienna, Austria.
  • Pflug FG; Center for Integrative Bioinformatics Vienna (CIBIV), Max F Perutz Laboratories (MFPL), University of Vienna, Medical University Vienna, Vienna, Austria.
  • Stirnberg A; Gregor Mendel Institute (GMI), Austrian Academy of Sciences, Vienna BioCenter (VBC), Vienna, Austria.
  • Ehrlinger K; Gregor Mendel Institute (GMI), Austrian Academy of Sciences, Vienna BioCenter (VBC), Vienna, Austria.
  • von Haeseler A; Center for Integrative Bioinformatics Vienna (CIBIV), Max F Perutz Laboratories (MFPL), University of Vienna, Medical University Vienna, Vienna, Austria.
  • Djamei A; Bioinformatics and Computational Biology, Faculty of Computer Science, University of Vienna, Vienna, Austria.
PLoS Biol ; 16(4): e2005129, 2018 04.
Article en En | MEDLINE | ID: mdl-29684023
Large-scale insertional mutagenesis screens can be powerful genome-wide tools if they are streamlined with efficient downstream analysis, which is a serious bottleneck in complex biological systems. A major impediment to the success of next-generation sequencing (NGS)-based screens for virulence factors is that the genetic material of pathogens is often underrepresented within the eukaryotic host, making detection extremely challenging. We therefore established insertion Pool-Sequencing (iPool-Seq) on maize infected with the biotrophic fungus U. maydis. iPool-Seq features tagmentation, unique molecular barcodes, and affinity purification of pathogen insertion mutant DNA from in vivo-infected tissues. In a proof of concept using iPool-Seq, we identified 28 virulence factors, including 23 that were previously uncharacterized, from an initial pool of 195 candidate effector mutants. Because of its sensitivity and quantitative nature, iPool-Seq can be applied to any insertional mutagenesis library and is especially suitable for genetically complex setups like pooled infections of eukaryotic hosts.
Asunto(s)
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Ustilago / Mutagénesis Insercional / Genoma Fúngico / Zea mays / Factores de Virulencia / Secuenciación de Nucleótidos de Alto Rendimiento Tipo de estudio: Prognostic_studies Idioma: En Revista: PLoS Biol Asunto de la revista: BIOLOGIA Año: 2018 Tipo del documento: Article País de afiliación: Austria Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Ustilago / Mutagénesis Insercional / Genoma Fúngico / Zea mays / Factores de Virulencia / Secuenciación de Nucleótidos de Alto Rendimiento Tipo de estudio: Prognostic_studies Idioma: En Revista: PLoS Biol Asunto de la revista: BIOLOGIA Año: 2018 Tipo del documento: Article País de afiliación: Austria Pais de publicación: Estados Unidos