New selectable marker/auxotrophic host strain combinations for molecular genetic manipulation of Pichia pastoris

We describe the isolation and characterization of three new biosynthetic genes-ARG4, ADE1, and URA3-from the methylotrophic yeast Pichia pastoris. The predicted products of the genes share significant sequence similarity to their Saccharomyces cerevisiae counterparts, namely argininosuccinate lyase,...

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Bibliographic Details
Published in:Gene 2001-01, Vol.263 (1-2), p.159-169
Main Authors: Lin Cereghino, G P, Lin Cereghino, J, Sunga, A J, Johnson, M A, Lim, M, Gleeson, M A, Cregg, J M
Format: Article
Language:English
Subjects:
ADE1 gene
Amino Acid Sequence
ARG4 gene
argininosuccinate lyase
Argininosuccinate Lyase - genetics
Cloning, Molecular
DNA, Fungal - chemistry
DNA, Fungal - genetics
Fungal Proteins - genetics
Fungal Proteins - metabolism
Genes, Fungal - genetics
Genetic Complementation Test
Genetic Markers
HIS4 gene
Methanol - metabolism
Molecular Sequence Data
Mutation
orotidine-5'-phosphate decarboxylase
Orotidine-5'-Phosphate Decarboxylase - genetics
Peptide Synthases - genetics
Pichia - enzymology
Pichia - genetics
Pichia pastoris
Plasmids - genetics
PR-aminoimidazolesuccinocarboxamide synthase
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Saccharomyces cerevisiae - enzymology
Saccharomyces cerevisiae - genetics
Sequence Analysis, DNA
Sequence Homology, Amino Acid
shuttle vectors
URA3 gene
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Summary:We describe the isolation and characterization of three new biosynthetic genes-ARG4, ADE1, and URA3-from the methylotrophic yeast Pichia pastoris. The predicted products of the genes share significant sequence similarity to their Saccharomyces cerevisiae counterparts, namely argininosuccinate lyase, PR-aminoimidazolesuccinocarboxamide synthase, and orotidine-5'-phosphate decarboxylase, respectively. Along with the previously described HIS4 gene, each gene was incorporated as the yeast selectable marker into a set of shuttle vectors designed to express foreign genes in P. pastoris. In addition, we have constructed a series of host strains containing all possible combinations of ade1, arg4, his4, and ura3 auxotrophies to be used with these new vectors.
ISSN:0378-1119
DOI:10.1016/s0378-1119(00)00576-x