Insertional mutagenesis in yeasts using T-DNA from Agrobacterium tumefaciens

Yeast
Paul BundockPaul J J Hooykaas

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.

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Citations

May 15, 2009·Plant Physiology·Stanton B Gelvin
Apr 13, 2004·Eukaryotic Cell·Alexander IdnurmJoseph Heitman
Jul 9, 2004·Genome Biology·Bart Scherens, Andre Goffeau
Mar 27, 2010·Annual Review of Phytopathology·Stanton B Gelvin
Sep 14, 2007·Molecular Microbiology·Jaehyuk ChoiYong-Hwan Lee
Oct 14, 2006·Molecular Microbiology·Christopher L MarionWilliam E Goldman
May 1, 2008·Microbial Biotechnology·Minna KemppainenAlejandro G Pardo
Jul 12, 2002·Yeast
Aug 17, 2005·Molecular Microbiology·Felicia J WaltonJoseph Heitman
Jun 5, 2007·Fungal Genetics and Biology : FG & B·Yan MengMark L Farman
Jun 30, 2007·Fungal Genetics and Biology : FG & B·Melania F BettsMarc J Orbach
May 13, 2005·Current Genetics·Caroline B MichielseArthur F J Ram

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