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Yeast
Article . 1999 . Peer-reviewed
License: Wiley TDM
Data sources: Crossref
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DIGITAL.CSIC
Article . 2013 . Peer-reviewed
Data sources: DIGITAL.CSIC
Yeast
Article . 1999
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Generation of null alleles for the functional analysis of six genes from the right arm ofSaccharomyces cerevisiae chromosome II

Authors: Dueñas, Encarnación; Vázquez de Aldana, Carlos R.; Cos, Teresa; Castro, Carmen; Valdivieso, María Henar;

Generation of null alleles for the functional analysis of six genes from the right arm ofSaccharomyces cerevisiae chromosome II

Abstract

Using PCR-ligated long flanking homology cassettes, null alleles of six open reading frames (ORFs) from chromosome II have been created in Saccharomyces cerevisiae. Deletants were constructed in three genetic backgrounds: FY1679, W303 and CEN.PK2. Tetrad analysis of heterozygous deletants revealed that none of the ORFs is essential for vegetative growth. Basic phenotypic analysis of haploid deletants showed that deletion of the YBR283c ORF causes a slight growth defect at 30 degrees C and 37 degrees C on glycerol-complete, glucose-complete, and glucose-minimal media only in the FY1679 and W303 backgrounds. Transformation of these deletants with the corresponding cognate gene in a centromeric plasmid complements the defects. Deletion of the YBR287w ORF leads to poor growth on glucose-minimal medium at 15 degrees C in the FY 1679 background. None of the six ORFs seems to be involved in mating or sporulation.

Keywords

Genes, Fungal, Restriction Mapping, Saccharomyces cerevisiae, Culture Media, Open Reading Frames, Phenotype, Transformation, Genetic, Chromosomes, Fungal, Alleles, Gene Deletion, Plasmids

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