Construction and high-throughput phenotypic screening ofZymoseptoria tritici over-expression strains.

摘要: Targeted gene deletion has been instrumental in elucidating many aspects of Zymoseptoria tritici pathogenicity. Gene over-expression is a complementary approach that amenable to rapid strain construction and high-throughput screening, which not exploited analyze Z. tritici, largely due lack available techniques. Here we exploit the Gateway® cloning technology for vectors improved homologous integration efficiency Δku70 build pilot library encompassing 32 genes encoding putative DNA binding proteins, GTPases or kinases. We developed protocol using Rotor-HDA robot reproducible cell pinning vitro screening. This screen identified an demonstrated marked reduction hyphal production relative isogenic progenitor. study provides generation libraries technique functional genomic screening this important pathogen.