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Generation of null alleles for the functional analysis of six genes from the right arm of Saccharomyces cerevisiae chromosome II
Author(s) -
Dueñas Encarnacion,
Vazquez De Aldana Carlos R.,
De Cos Teresa,
Castro Carmen,
Valdieso M. Henar
Publication year - 1999
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(199905)15:7<615::aid-yea385>3.0.co;2-x
Subject(s) - orfs , biology , genetics , open reading frame , null allele , gene , saccharomyces cerevisiae , plasmid , allele , homology (biology) , chromosome , genetic analysis , peptide sequence
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°C and 37°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°C in the FY1679 background. None of the six ORFs seems to be involved in mating or sporulation. Copyright © 1999 John Wiley & Sons, Ltd.