title: Insertional mutagenesis in yeasts using T-DNA from Agrobacterium tumefaciens.
abstract: Insertional mutagenesis is a powerful tool for the isolation of novel mutations. The gene delivery system of the bacterium Agrobacterium tumefaciens, which mediates transfer not only to plants but also to yeasts and fungi, could be exploited to generate collections of yeasts containing insertional mutations if there were no bias towards particular integration sites, as is the case in plants. To test this, we have analysed a small collection of Saccharomyces cerevisiae strains with T-DNA copies integrated in the S. cerevisiae genome. The position of 54 of these T-DNAs was determined. The T-DNA showed no clear preference for certain DNA sequences or genomic regions. We have isolated insertions in the coding regions of the genes YGR125w, YDR250c, YGR141w, YGR045c, YPL017c, YGR040w, YDL052c, YJL148w, YCL033c, YFL061w, YJR033c, YDR175c and YLR309c confirming that these genes are non-essential for S. cerevisiae haploid growth on minimal medium. Given the advantages of T-DNA, we propose its use as an ideal mobile DNA element for insertional mutagenesis in yeasts.
authors.affiliation: Institute for Molecular Plant Sciences, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands. bundock@rulbim.leidenuniv.nl
authors.forename: Paul
authors.initials: P
authors.lastname: Bundock
authors.forename: Haico
authors.initials: H
authors.lastname: van Attikum
authors.forename: Amke
authors.initials: A
authors.lastname: den Dulk-Ras
authors.forename: Paul J J
authors.initials: PJ
authors.lastname: Hooykaas
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D005800:Genes, Fungal; D006239:Haplotypes; D016254:Mutagenesis, Insertional; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development
chemical_list: D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; C036483:T-DNA