Integration of an insertion-type transferred DNA vector from Agrobacterium tumefaciens into the Saccharomyces cerevisiae genome by gap repair.
Recently, it was shown that Agrobacterium tumefaciens can transfer transferred DNA (T-DNA) to Saccharomyces cerevisiae and that this T-DNA, when used as a replacement vector, is integrated via homologous recombination into the yeast genome. To test whether T-DNA can be a suitable substrate for integration via the gap repair mechanism as well, a model system developed for detection of homologous recombination events in plants was transferred to S. cerevisiae. Analysis of the yeast transformants revealed that an insertion type T-DNA vector can indeed be integrated via gap repair. Interestingly, the transformation frequency and the type of recombination events turned out to depend strongly on the orientation of the insert between the borders in such an insertion type T-DNA vector.
Risseeuw
E
E
Clusius Laboratory, Institute of Molecular Plant Sciences, Leiden University, The Netherlands.
Franke-van Dijk
M E
ME
Hooykaas
P J
PJ
D016960:Agrobacterium tumefaciens; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers; D004260:DNA Repair; D004269:DNA, Bacterial / Q000378:metabolism*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D016681:Genome, Fungal; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011995:Recombination, Genetic; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D017931:DNA Primers; D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; C036483:T-DNA