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Contribution to Book
Competent Cell Preparation and Transformation of Pichia pastoris
Yeast Metabolic Engineering
  • Joan Lin-Cereghino, University of California, San Diego
  • Christopher A Naranjo, University of the Pacific
  • Geoff Lin-Cereghino, University of the Pacific
Document Type
Contribution to Book
Department
Biological Sciences
Editor(s)
Valeria Mapelli & Maurizio Bettiga
Description

During the past three decades, the methylotrophic yeast Pichia pastoris (recently reclassified as Komagataella phaffii) has gained widespread acceptance as a system of choice for heterologous protein expression. One of the reasons that this yeast is used so frequently is the simplicity of techniques required for its molecular genetic manipulation. There are several different protocols available for introducing DNA into P. pastoris using electroporation or heat shock. We describe here a shortened protocol for cell preparation and transformation that works reliably with either prototrophic markers or antibiotic selection in this host. This procedure utilizes the most efficient portions of the electroporation and heat-shock transformation protocols to yield a method that is both time-saving and effective.

Find in WorldCat
https://worldcat.org/title/1334605192
ISBN
978-1-0716-2399-2
DOI
10.1007/978-1-0716-2399-2_7
Publication Date
7-4-2022
Publisher
Humana Press
Keywords
  • Transformation,
  • Electroporation,
  • Recombinant proteins,
  • Protein expression,
  • Pichia pastoris
Disciplines
Comments

Part of the book series: Methods in Molecular Biology (MIMB, volume 2513)

https://link.springer.com/protocol/10.1007/978-1-0716-2399-2_7
Citation Information
Joan Lin-Cereghino, Christopher A Naranjo and Geoff Lin-Cereghino. "Competent Cell Preparation and Transformation of Pichia pastoris" New YorkYeast Metabolic Engineering (2022) p. 113 - 120
Available at: http://works.bepress.com/joan-lin-cereghino/83/