Agrobacterium tumefaciens-mediated transformation of yeast

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 93; no. 4; pp. 1613 - 1618
• Main Authors: Piers, K.L. (Red Deer College, Red Deer, Alberta, Canada.), Heath, J.D, Liang, X.Y, Stephens, K.M, Nester, E.W
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 20.02.1996; National Acad Sciences; National Academy of Sciences
• Subjects: ADN; AGROBACTERIUM TUMEFACIENS; Agrobacterium tumefaciens - genetics; Agrobacterium tumefaciens - pathogenicity; Bacteria; Bacterial Proteins - genetics; Bacterial Proteins - physiology; Coculture techniques; Deoxyribonucleic acid; DNA; DNA, Bacterial - genetics; DNA, Fungal - genetics; DNA, Recombinant - genetics; Flowers & plants; GENE; GENES; Genes, Bacterial; Genetic Vectors; Integration host factors; Microbiology; Molecular biology; Plant cells; PLASMIDE; PLASMIDIOS; Plasmids; SACCHAROMYCES CEREVISIAE; Saccharomyces cerevisiae - genetics; Sequence Deletion; Telomere - genetics; Transfection; TRANSFORMACION GENETICA; TRANSFORMATION GENETIQUE; Virulence; Virulence - genetics; Virulence Factors; Yeast; Yeasts
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.93.4.1613

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.