Generation and analysis of a barcode-tagged insertion mutant library in the fission yeast Schizosaccharomyces pombe.

BMC Genomics
Bo-Ruei ChenKurt W Runge

Abstract

Barcodes are unique DNA sequence tags that can be used to specifically label individual mutants. The barcode-tagged open reading frame (ORF) haploid deletion mutant collections in the budding yeast Saccharomyces cerevisiae and the fission yeast Schizosaccharomyces pombe allow for high-throughput mutant phenotyping because the relative growth of mutants in a population can be determined by monitoring the proportions of their associated barcodes. While these mutant collections have greatly facilitated genome-wide studies, mutations in essential genes are not present, and the roles of these genes are not as easily studied. To further support genome-scale research in S. pombe, we generated a barcode-tagged fission yeast insertion mutant library that has the potential of generating viable mutations in both essential and non-essential genes and can be easily analyzed using standard molecular biological techniques. An insertion vector containing a selectable ura4+ marker and a random barcode was used to generate a collection of 10,000 fission yeast insertion mutants stored individually in 384-well plates and as six pools of mixed mutants. Individual barcodes are flanked by Sfi I recognition sites and can be oligomerized in a unique or...Continue Reading

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Citations

Nov 3, 2015·Critical Reviews in Biochemistry and Molecular Biology·Charalampos Rallis, Jürg Bähler
Nov 7, 2012·Current Opinion in Cell Biology·Olivier HachetSophie G Martin
May 23, 2014·Frontiers in Genetics·Brandon D Gaytán, Chris D Vulpe
Sep 30, 2020·The Journal of Microbiology·Kwang-Woo JungYong-Sun Bahn
Aug 3, 2017·Cold Spring Harbor Protocols·Karl Ekwall, Geneviève Thon
Mar 18, 2021·Briefings in Functional Genomics·Luc Legon, Charalampos Rallis

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Methods Mentioned

BETA
PCR
electrophoresis

Software Mentioned

AmiGO

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