Construction of fission yeast vectors with a novel selection strategy that allows their use in wild-type fission yeasts.
Novel vectors that use the Pichia pastoris INO1 gene as a selectable marker and exploit the natural inositol auxotrophy of the fission yeast are described. These plasmids also contained other features desirable in a plasmid cloning vector. These plasmids were evaluated in other species of Schizosaccharomyces and found to replicate autonomously in another variety of S. pombe, S. pombe var. malidevorans. These plasmids can be used for transformation of any wild-type S. pombe strain without the need for selection by induced auxotrophic mutations, or by selection by drug resistance markers, and should greatly assist genetic and molecular manipulations in these yeasts.
Ingavale
S S
SS
Institute of Microbial Technology, Sector 39-A, Chandigarh- 160 036, India.
Sharma
K G
KG
Bachhawat
A K
AK
D003001:Cloning, Molecular; D003470:Culture Media; D005800:Genes, Fungal; D005819:Genetic Markers; D005822:Genetic Vectors; D007294:Inositol / Q000378:metabolism*; D007296:Myo-Inositol-1-Phosphate Synthase / Q000235:genetics* / Q000378:metabolism; D010843:Pichia / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
D003470:Culture Media; D005819:Genetic Markers; D007294:Inositol; D007296:Myo-Inositol-1-Phosphate Synthase