A vector system for genomic FLAG epitope-tagging in Schizosaccharomyces pombe

Biotechnology Journal
Chiaki NoguchiEishi Noguchi

Abstract

The fission yeast Schizosaccharomyces pombe is a popular model organism to study various cellular processes, although research tools available for S. pombe are relatively inadequate. To facilitate genetic and biochemical investigation in S. pombe, we report here a system of vectors for genomic FLAG epitope-tagging. These vectors enable us to amplify gene-targeting fragments for integration into specific loci of the S. pombe genome. All vectors in this report were designed to express FLAG epitope-tagged proteins from their endogenous genomic loci. Vectors for N-terminal FLAG epitope-tagging allow us to control protein expression levels using the wild-type nmt1 promoter, its weaker derivatives, and the urg1 promoter. These vectors are available with various antibiotic markers including kanMX6, hphMX6, natMX6 and bleMX6, and the his3(+) marker. Vectors for C-terminal FLAG epitope-tagging were designed to express FLAG-fusion proteins under the control of their native promoters at their own genomic loci, allowing us to characterize protein functions under physiological conditions. These vectors are available with kanMX6, hphMX6, nat-MX6 and bleMX6 markers. The series of vectors described in this report should prove useful for protei...Continue Reading

References

Jan 1, 1991·Methods in Enzymology·S MorenoP Nurse
Sep 22, 1999·Molecular and Cellular Biology·Y OgawaH Masukata
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Citations

Nov 9, 2011·Molecular and Cellular Biology·Lynda M GroocockMichael N Boddy
Nov 13, 2013·BioTechniques·Mariana C GadaletaEishi Noguchi
Mar 19, 2016·PLoS Genetics·Mariana C GadaletaEishi Noguchi
Aug 9, 2017·Molecular and Cellular Biology·Michael C ReubensPaul Russell
Dec 14, 2018·The Journal of Biological Chemistry·Christopher J ZysnarskiMara C Duncan
Mar 11, 2020·Frontiers in Microbiology·Lianfang WangEleftherios Mylonakis
Feb 2, 2019·Nature Communications·Yu-Hua LoRobin E Stanley
Jul 10, 2019·Current Biology : CB·Su Ling LeongKenneth E Sawin
Jan 23, 2021·Life Science Alliance·Tianyao XiaoAdam L Hughes
Feb 5, 2021·Nature Communications·Vedrana AndricMathieu Rougemaille

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