Prion Transfection of Yeast.
Transfection of yeast with amyloid filaments, made from recombinant protein or prepared from extracts of cells infected with a prion, has become an important method in characterizing yeast prions. Here, we describe a method for transmission of [URE3] with Ure2p amyloid that is based on a previously published protocol for transfection with Sup35p filaments to make cells [PSI+]. This method may be used for other prions by changing just the amyloid source, host strain, and plating medium.
Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892-0830.
Herman K
HK
Edskes
Department of Pharmacology, Uniformed Services University for the Health Sciences, Bethesda, Maryland 20814.
Dmitry
D
Kryndushkin
Department of Pharmacology, Uniformed Services University for the Health Sciences, Bethesda, Maryland 20814.
Frank
F
Shewmaker
Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892-0830; wickner@helix.nih.gov.
Reed B
RB
Wickner
D005979:Glutathione Peroxidase / Q000235:genetics* / Q000378:metabolism*; D010641:Phenotype; D000072002:Prion Proteins / Q000378:metabolism*; D011328:Prions / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
D000072002:Prion Proteins; D011328:Prions; D029701:Saccharomyces cerevisiae Proteins; D005979:Glutathione Peroxidase; C067020:URE2 protein, S cerevisiae