Using the DHFR heat-inducible degron for protein inactivation in Schizosaccharomyces pombe.
Inactivating a specific protein in vivo can yield important information about its function. One strategy previously developed in Saccharomyces cerevisiae by the Varshavsky group involves fusing a degron, derived from mouse dihydrofolate reductase, to the N-terminus of the target protein, which thereby confers temperature-sensitive degradation at the restrictive temperature. We describe here the application of this technique in the fission yeast, Schizosaccharomyces pombe.
Kearsey
Stephen E
SE
Department of Zoology, University of Oxford, Oxford, UK.
Gregan
Juraj
J
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D005822:Genetic Vectors; D006358:Hot Temperature; D066298:In Vitro Techniques; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D014170:Transformation, Genetic
D017931:DNA Primers; D004274:DNA, Recombinant; D011993:Recombinant Fusion Proteins; D029702:Schizosaccharomyces pombe Proteins; D013762:Tetrahydrofolate Dehydrogenase