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Disruption and basic phenotypic analysis of six novel genes from the right arm of chromosome XII of Saccharomyces cerevisiae
Author(s) -
Watson Martin D.
Publication year - 2001
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/yea.704
Subject(s) - orfs , biology , genetics , saccharomyces cerevisiae , open reading frame , plasmid , gene , terminator (solar) , homology (biology) , mutant , mating type , peptide sequence , ionosphere , physics , astronomy
Six open reading frames (ORFs) of unknown function from the right arm of Saccharomyces cerevisiae chromosome XII were deleted in two genetic backgrounds by disruption cassettes with regions of short flanking homology. This work was carried out within the framework of the EUROFAN consortium. The SFH disruption cassettes, obtained by PCR, were made by amplification of the kanMX marker module with oligonucleotides containing approximately 40 bp of homology to either the promoter or translation terminator regions of the relevant ORF. Transformants resistant to geneticin (G418) were selected. The SFH disruption cassettes were cloned into a bacterial vector. Each cognate gene was also cloned into a yeast centromeric plasmid. Sporulation and tetrad analysis of the disrupted heterozygous strains revealed that ORF YLR153c (now known as ACS2 ) is essential. Basic phenotypic analysis was performed on haploid deletants of both mating types of the five non‐essential ORFs, YLR082c (now known as SRL2 ), YLR149c, YLR151c, YLR152c and YLR154c. Plate growth tests on different media at 15°C, 30°C and 37°C did not reveal any significant differences between parental and mutant cells. Mating and sporulation efficiencies were not affected in any of the viable disruptants as compared to wild‐type cells. Copyright © 2001 John Wiley & Sons, Ltd.