NR AVDS
AU King,C.Y.; Wang,H.L.; Chang,H.Y.
TI Transformation of yeast by infectious prion particles
QU Methods (San Diego, Calif.) 2006 May; 39(1): 68-71
PT journal article
AB We present methods to prepare infectious Sup35 protein aggregates and use them for genetic transformation of yeast. The protein aggregates are prepared from bacterially expressed recombinant protein, which is converted to amyloid fibers by extended incubation or by nucleated growth using yeast prion particles as seeds. The aggregates are introduced into yeast by a modified spheroplast transformation protocol. The phenotype of the yeast transformants is further characterized by robust prion strain typing methods. The methodology can be used to introduce different [PSI(+)] particles to many laboratory yeast genetic backgrounds. It can be adapted for applications in other yeast prion systems as well.
MH Amyloid/chemistry; Cell Fusion/methods; Escherichia coli/genetics; Green Fluorescent Proteins/chemistry/genetics/metabolism; Mutation, Missense/genetics; Pigmentation; Prions/chemistry/*genetics/metabolism; Recombinant Fusion Proteins/chemistry/genetics/metabolism; Saccharomyces cerevisiae/cytology/*genetics; Saccharomyces cerevisiae Proteins/chemistry/*genetics/metabolism; Spheroplasts/genetics; Transformation, Genetic
AD Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan. cking@imb.sinica.edu.tw
SP englisch
PO USA