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Gene disruption and basic phenotypic analysis of nine novel yeast genes from chromosome XIV
Author(s) -
Capozzo Chiara,
Sartorello Francesca,
Dal Pero Francesca,
D'Angelo Michela,
Vezzi Alessandro,
Campanaro Stefano,
Valle Giorgio
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 - 10.1002/1097-0061(20000915)16:12<1089::aid-yea600>3.0.co;2-g
Subject(s) - biology , orfs , gene , homology (biology) , genetics , phenotype , ploidy , homologous chromosome , tetrad , saccharomyces cerevisiae , chromosome , yeast , microbiology and biotechnology , open reading frame , peptide sequence
In this work, we describe the disruption of nine ORFs of S. cerevisiae (YNL123w, YNL119w, YNL115c, YNL108c, YNL110c, YNL124w, YNL233w, YNL232w and YNL231c) in two genetic backgrounds: FY1679 and CEN.PK2. For the construction of the deletant strains, we used the strategy of short flanking homology (SFH) PCR. The SFH‐deletion cassette was made by PCR amplification of the KanMX4 module with primers containing a 5′ region of 40 bases homologous to the target yeast gene and with a 3′ region of 20 bases homologous to pFA6a–KanMX4 MCS. Sporulation and tetrad analysis of heterozygous deletants revealed that YNL110c, YNL124w and YNL232w are essential genes. The subcellular localization of the protein encoded by the essential gene YNL110c was investigated using the green fluorescent protein (GFP) approach, revealing a nuclear pattern. Basic phenotypic analysis of the non‐essential genes revealed that the growth of ynl119w Δ haploid cells was severely affected at 37°C in N3 medium, indicating that this gene is required at high temperatures with glycerol as a non‐fermentable substrate. The ynl233w Δ haploid cells also showed a particular phenotype under light microscopy and were studied in detail in a separate work. Copyright © 2000 John Wiley & Sons, Ltd.