Selecting Schizosaccharomyces pombe Diploids

Cold Spring Harb Protoc. 2017 Jul 5;2017(7):pdb.prot091702. doi: 10.1101/pdb.prot091702.

Abstract

Here we describe procedures for the selection of diploid Schizosaccharomyces pombeade6-M210/ade6-M216 heteroallelic complementation is widely used to select for Ade+ diploids. Such diploids will readily sporulate when starved of nitrogen. For some investigations, stable diploids are preferable (e.g., for genetic complementation tests), and in these cases mating an h- strain with an h90 mat2-Pi-102 strain can be used to prevent sporulation. When ade6-M210/ade6-M216 mutations impact on, or show synthetic interactions with, the gene of interest, two different auxotrophic markers can be used to select complementation.

MeSH terms

  • Culture Media / chemistry
  • Diploidy*
  • Genes, Fungal
  • Genetics, Microbial / methods*
  • Microbiological Techniques / methods*
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces / growth & development
  • Schizosaccharomyces / isolation & purification*
  • Selection, Genetic*

Substances

  • Culture Media