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Disruption and functional analysis of six ORFs on chromosome IV: YDR013w, YDR014w, YDR015c, YDR018c, YDR020c, YDR021w ( FAL1 )
Author(s) -
Dardalhon Michèle,
MagañaSchwencke Nieve,
Averbeck Dietrich
Publication year - 2000
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(200002)16:3<267::aid-yea522>3.0.co;2-n
Subject(s) - orfs , biology , genetics , open reading frame , ploidy , gene , locus (genetics) , mutant , chromosome , phenotype , homologous chromosome , genetic analysis , peptide sequence
The disruption of six novel yeast genes has been realized in two genetic backgrounds. Six open reading frames (ORFs) from chromosome IV, YDR013w, YDR014w, YDR015c, YDR018c, YDR020c and YDR021w, were disrupted using the KanMX4 marker and PCR‐targeting with long flanking regions homologous (LFH) to the target locus. The deletants were verified at the molecular level, using PCR and Southern analysis. Sporulation and tetrad analysis revealed that ORFs YDR013w and YDR021w (also known as FAL1 ) are essential genes. Microscopical observations showed that ydr013w Δ haploid cells were blocked after one or two cell cycles and presented heterogeneous bud sizes. The ydr021w Δ haploid cells gave rise to microcolonies of about 20 cells. The other four ORFs are non‐essential. Basic phenotypic analysis of the non‐lethal deletant strains did not reveal any significant differences in cell morphology, growth on different media and temperatures, sporulation and mating efficiency between parental and mutant strains in the FY1679 background. Copyright © 2000 John Wiley & Sons, Ltd.