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Deletion of six open reading frames from the left arm of chromosome IV ofSaccharomyces cerevisiae

✍ Scribed by Tuller, Gabriele; Prein, Birgit; Jandrositz, Anita; Daum, Günther; Kohlwein, Sepp D.

Publisher
John Wiley and Sons
Year
1999
Tongue
English
Weight
336 KB
Volume
15
Category
Article
ISSN
0749-503X

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✦ Synopsis

The construction of six deletion mutants of Saccharomyces cerevisiae and their basic phenotypic characterization are described. Open reading frames YDL148c, YDL109c, YDL021w, YDL019c, YDL018c and YDL015c from the left arm of chromosome IV were deleted using a polymerase chain reaction (PCR)-based disruption technique, introducing the kanMX4 resistance marker into the respective genes. Gene replacement cassettes (pYORCs) for use in other strain backgrounds were cloned by PCR using DNA templates from haploid or diploid deletion mutants, and inserted into episomal plasmids. Cognate clones of all six ORFs were obtained by gap repair. Deletions were carried out in diploid cells and, after sporulation, yielded four viable spores for clones disrupted in YDL109c, YDL021w, YDL019c and YDL018c. Spores harbouring disruptions in ORFs YDL148c and YDL015c germinated but underwent only a few divisions before ceasing growth, suggesting that the respective genes are essential for vegetative growth on YPD complete media. The other deletion mutants grew like wild-type at different temperatures and on different carbon sources. A brief computational analysis of the six ORFs studied in this work is presented.

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