title: Development of episomal vectors carrying a nourseothricin-resistance marker for use in minimal media for Schizosaccharomyces pombe.
abstract: In the post-genomic era, an immediate challenge is to assign biological functions to novel proteins encoded by the genome. This challenge requires the use of a simple organism as a genetic tool and a range of new high-throughput techniques. Schizosacchromyces pombe is a powerful model organism used to investigate disease-related genes and provides useful tools for the functional analysis of heterologous genes. To expand the current array of experimental tools, we constructed two series of Sz. pombe expression vectors, i.e. general and Gateway vectors, containing nourseothricin-resistance markers. Vectors carrying nourseothricin-resistance markers possess advantages in that they do not limit the parental strains with auxotrophic mutations with respect to availability for use in clone selection and can be used together with vectors carrying nutrient markers in minimal media. We modified the pSLF173, pSLF273 and pSLF373 vectors carrying a triple haemagglutinin epitope (3×HA) and an Ura4 marker. The vectors described here contain the nmt1 promoter with three different episomal expression strengths for proteins fused with 3×HA, EGFP or DsRed at the N-terminus. These vectors represent an important contribution to the genome-wide investigation of multiple heterologous genes and for functional and genetic analysis of novel human genes.
authors.affiliation: Genome Research Centre, KRIBB, Daejeon, 305-806, Republic of Korea.
authors.forename: Jiwon
authors.initials: J
authors.lastname: Ahn
authors.forename: Misun
authors.initials: M
authors.lastname: Won
authors.forename: Mi-Lang
authors.initials: ML
authors.lastname: Kyun
authors.forename: Yong Sung
authors.initials: YS
authors.lastname: Kim
authors.forename: Cho-Rock
authors.initials: CR
authors.lastname: Jung
authors.forename: Dong-Su
authors.initials: DS
authors.lastname: Im
authors.forename: Kyung-Bin
authors.initials: KB
authors.lastname: Song
authors.forename: Kyung-Sook
authors.initials: KS
authors.lastname: Chung
mesh_terms: D000123:Acetyltransferases / Q000235:genetics*; D003470:Culture Media; D025141:Drug Resistance, Fungal / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D005822:Genetic Vectors / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins; D012568:Schizosaccharomyces / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D013309:Streptothricins / Q000494:pharmacology*
chemical_list: D003470:Culture Media; D011993:Recombinant Fusion Proteins; D013309:Streptothricins; D000123:Acetyltransferases; C050473:nourseothricin acetyltransferase