Molecular biology. Transformation: introduction of DNA Selectable marker Spheroplasts, Li 2+ salts, electroporation Yeast plasmids are shuttle plasmids, amplification in E. coli, ori, b -lactamase Transformation with oligonucleotides, -> selection
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Transformation: introduction of DNA
Spheroplasts, Li2+ salts, electroporation
Yeast plasmids are shuttle plasmids, amplification in E. coli,
Transformation with oligonucleotides, -> selection
Transformation of mitochondria by particle bombardment
The YFG1 +gene is disrupted by transforming the strain with a linear fragment containing a URA3 selectable marker flanked by homologous sequences. The chromosomal segment is replaced by this URA3 containing fragment after integration by homologous recombination.
The URA3 marker introduced in the YFG1 locus, can be excised if URA3 is also flanked by direct repeats of DNA, preferably not originating from yeast. Homologous recombinants lack the URA3 marker and retain a single copy of the repeated DNA.
Single-step gene replacement of mutant alleles, such as yfg1-1 , can be carried out by first replacing the YFG1 gene by URA3 , transforming the strain with linear fragment encompassing the yfg1-1 mutation, and selecting transformants in which URA3 is replaced by yfg1-1.
Microarrays (DNA, oligo)
comparative, yeast (sensus stricto),...
Proteomics, MS, chips (120 kinases)
Metabolomics, flux of metabolites