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Direct selection of Pichia pastoris expression strains using new G418 resistance vectors
Author(s) -
LinCereghino Joan,
Hashimoto Matthew D.,
Moy Allison,
Castelo James,
Orazem Claire C.,
Kuo Peter,
Xiong See,
Gandhi Vishal,
Hatae Christopher T.,
Chan Alex,
LinCereghino Geoff P.
Publication year - 2008
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.1587
Subject(s) - pichia pastoris , selectable marker , biology , transformation (genetics) , heterologous , multiple cloning site , genbank , cloning (programming) , expression vector , gene , genetics , computational biology , vector (molecular biology) , heterologous expression , accession number (library science) , recombinant dna , computer science , programming language
The methylotrophic yeast, Pichia pastoris, is widely used as a host organism for the expression of heterologous proteins. Currently, the Zeocin and blasticidin resistance genes are the only dominant selectable markers that can be used for primary selection of transformants. In this report we describe new expression vectors that can be used to select directly for P. pastoris transformants using G418 resistance conferred by a modified Tn903kan r gene. Compared to other dominant markers, this system is more economical and offers a higher transformation efficiency, due to the small sizes of the cloning vectors, pKAN B and pKANα B (GenBank Accession Nos EU285585 and EU285586, respectively). Additionally, multicopy transformants can be generated using these new vectors. Copyright © 2008 John Wiley & Sons, Ltd.