Competent Cell Preparation and Transformation of Pichia pastoris
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Document Type
Contribution to Book
Department
Biological Sciences
Book Title
Yeast Metabolic Engineering
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.
Buy Link
https://link.springer.com/protocol/10.1007/978-1-0716-2399-2_7
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
City
New York
First Page
113
Last Page
120
Keywords
Transformation, Electroporation, Recombinant proteins, Protein expression, Pichia pastoris
Disciplines
Life Sciences
Recommended Citation
Lin-Cereghino, J.,
Naranjo, C. A.,
&
Lin-Cereghino, G.
(2022).
Competent Cell Preparation and Transformation of Pichia pastoris.
In Valeria Mapelli & Maurizio Bettiga (Eds.), Yeast Metabolic Engineering (113–120). New York: Humana Press
https://scholarlycommons.pacific.edu/cop-facbooks/231
https://link.springer.com/protocol/10.1007/978-1-0716-2399-2_7
Comments
Part of the book series: Methods in Molecular Biology (MIMB, volume 2513)