Agrobacterium tumefaciens-mediated transformation of yeast.
Agrobacterium tumefaciens transfers a piece of its Ti plasmid DNA (transferred DNA or T-DNA) into plant cells during crown gall tumorigenesis. A. tumefaciens can transfer its T-DNA to a wide variety of hosts, including both dicotyledonous and monocotyledonous plants. We show that the host range of A. tumefaciens can be extended to include Saccharomyces cerevisiae. Additionally, we demonstrate that while T-DNA transfer into S. cerevisiae is very similar to T-DNA transfer into plants, the requirements are not entirely conserved. The Ti plasmid-encoded vir genes of A. tumefaciens that are required for T-DNA transfer into plants are also required for T-DNA transfer into S. cerevisiae, as is vir gene induction. However, mutations in the chromosomal virulence genes of A. tumefaciens involved in attachment to plant cells have no effect on the efficiency of T-DNA transfer into S. cerevisiae. We also demonstrate that transformation efficiency is improved 500-fold by the addition of yeast telomeric sequences within the T-DNA sequence.
Piers
K L
KL
Department of Microbiology, University of Washington, Seattle, 98195, USA.
Heath
J D
JD
Liang
X
X
Stephens
K M
KM
Nester
E W
EW
D016960:Agrobacterium tumefaciens / Q000235:genetics* / Q000472:pathogenicity; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D004269:DNA, Bacterial / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics*; D005798:Genes, Bacterial; D005822:Genetic Vectors; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D017384:Sequence Deletion; D016615:Telomere / Q000235:genetics*; D014162:Transfection; D014774:Virulence / Q000235:genetics; D037521:Virulence Factors
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004271:DNA, Fungal; D004274:DNA, Recombinant; D037521:Virulence Factors