Generation of disruption cassettes in vivo using a PCR product and Saccharomyces cerevisiae.
A method to obtain disruption cassettes based on the homologous recombination in Saccharomyces cerevisiae is described. The disruption marker is amplified by PCR using oligonucleotides containing 50 bp homologous to the disruptable gene and 20 bp from the marker. The PCR product is cotransformed into yeast with a plasmid containing the gene. After recombination, a plasmid that carries the disruption cassette for the gene is produced.
Zaragoza
Oscar
O
Instituto de Investigaciones Biomédicas Alberto Sols CSIC-UAM, C/Arturo Duperier 4, Madrid 28029, Spain. ozaragoz@aecom.yu.edu
D001483:Base Sequence; D005819:Genetic Markers; D016681:Genome, Fungal; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D005819:Genetic Markers