Direct selection of Pichia pastoris expression strains using new G418 resistance vectors.
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 pKANalpha B (GenBank Accession Nos EU285585 and EU285586, respectively). Additionally, multicopy transformants can be generated using these new vectors.
Lin-Cereghino
Joan
J
Department of Biological Sciences, University of the Pacific, Stockton, CA 95211-0197, USA.
Hashimoto
Matthew D
MD
Moy
Allison
A
Castelo
James
J
Orazem
Claire C
CC
Kuo
Peter
P
Xiong
See
S
Gandhi
Vishal
V
Hatae
Christopher T
CT
Chan
Alex
A
Lin-Cereghino
Geoff P
GP
D003001:Cloning, Molecular; D018628:Gene Dosage; D005821:Genetic Techniques; D005822:Genetic Vectors; D005839:Gentamicins / Q000494:pharmacology*; D007612:Kanamycin / Q000494:pharmacology; D010843:Pichia / Q000187:drug effects / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012331:RNA, Fungal / Q000235:genetics; D012641:Selection, Genetic; D014170:Transformation, Genetic
D005839:Gentamicins; D012331:RNA, Fungal; D007612:Kanamycin; C010680:antibiotic G 418