Pichia protocols /

Bibliographic Details
Other Authors: Higgins, David R., Cregg, James M.
Format: Book
Language:English
Published: Totowa, N.J. : Humana Press, [1998]
Series:Methods in molecular biology (Clifton, N.J.) ; v. 103.
Subjects:
Table of Contents:
  • Introduction to Pichia pastoris
  • Classical genetic manipulation
  • Transformation
  • Small vectors for expression based on dominant drug resistance with direct multicopy selection
  • The generation of multicopy recombinant strains
  • Isolation of nucleic acids
  • Generation of pretease-deficient strains and their use in hererologous protein expression
  • Glycosylation profiling of heterologous proteins
  • High cell-density fermentation
  • Use of Pichia pastoris as a model eukaryotic system : peroxisome biogenesis
  • Secretion of recombinant human insulin-like growth factor I (IGF-I)
  • Secretion of scFv antibody fragments
  • Expression of tetanus toxin fragment C
  • Expression of EGF and HIV envelope glycoprotein
  • Expression of an integral membrane protein, the 5HT̐₅ receptor
  • Localization of the myc-tagged 5HT̐₅ receptor by immunogold staining of ultrathin sections