Methods of plate pexophagy monitoring and positive selection for ATG gene cloning in yeasts.
Methods for colony assay of peroxisomal oxidases in yeasts provide a convenient and fast approach for monitoring peroxisome status. They have been used in several laboratories for the isolation of yeast mutants deficient in selective autophagic peroxisome degradation (pexophagy), catabolite repression of peroxisomal enzymes or mutants deficient in oxidases themselves. In this chapter, protocols for monitoring peroxisomal alcohol oxidase and amine oxidase directly in yeast colonies and examples of their application for mutant isolation are described. These methods were successfully utilized in several methylotrophic yeasts and the alkane-utilizing yeast Yarrowia lipolytica.
Stasyk
Oleh V
OV
Institute of Cell Biology, NAS of Ukraine, Lviv, Ukraine.
Nazarko
Taras Y
TY
Sibirny
Andriy A
AA
D000429:Alcohol Oxidoreductases / Q000378:metabolism; D001343:Autophagy / Q000502:physiology*; D001681:Biological Assay / Q000379:methods*; D002374:Catalase / Q000378:metabolism; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D020675:Peroxisomes / Q000201:enzymology*; D010843:Pichia / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D020005:Propanols / Q000378:metabolism; D025062:Yarrowia / Q000166:cytology / Q000235:genetics / Q000378:metabolism
D005656:Fungal Proteins; D020005:Propanols; C006463:allyl alcohol; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D002374:Catalase