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Disruption and functional analysis of six ORFs on chromosome XII of Saccharomyces cerevisiae: YLR124w, YLR125w, YLR126c, YLR127c, YLR128w and YLR129w

✍ Scribed by Martin Willer; Matthieu Regnacq; Peter J. Reid; John R. Tyson; Wei Cui; Barrie M. Wilkinson; Colin J. Stirling


Publisher
John Wiley and Sons
Year
2000
Tongue
English
Weight
188 KB
Volume
16
Category
Article
ISSN
0749-503X

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✦ Synopsis


In the context of the EUROFAN programme, we report the deletion and functional analysis of six open reading frames (ORFs) on the right arm of chromosome XII of Saccharomyces cerevisiae. Using a PCR-based gene replacement strategy, we have systematically deleted individual ORFs and subjected the heterozygous diploids and haploid knockout strains to basic genetic and phenotypic characterization. Two ORFs, YLR127c and YLR129w, are essential for viability, whereas no growth phenotype could be detected following deletion of YLR124w, YLR125w, YLR126c or YLR128w. For each of the individual ORFs, a kanMX4 replacement cassette and the corresponding cognate wild-type gene were cloned into appropriate plasmids.


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Six novel Open Reading Frames (ORFs) located on the left arm of chromosome XII (YLL044w, YLL042c, YLL040c, YLL038c, YLL035w and YLL034c) have been analysed using short-Β―anking homology (SFH) gene replacement. Sporulation and tetrad analysis showed that YLL035w and YLL034c are essential for cell grow