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, 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.
Lin Cereghino
G P
GP
Department of Biochemistry and Molecular Biology, Oregon Graduate Institute of Science and Technology, 20000 N.W. Walker Road, Beaverton, OR 97006-8921, USA.
Lin Cereghino
J
J
Sunga
A J
AJ
Johnson
M A
MA
Lim
M
M
Gleeson
M A
MA
Cregg
J M
JM
D000595:Amino Acid Sequence; D001123:Argininosuccinate Lyase / Q000235:genetics; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D005800:Genes, Fungal / Q000235:genetics; D005816:Genetic Complementation Test; D005819:Genetic Markers; D000432:Methanol / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000235:genetics; D010453:Peptide Synthases / Q000235:genetics; D010843:Pichia / Q000201:enzymology / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D004271:DNA, Fungal; D005656:Fungal Proteins; D005819:Genetic Markers; D011993:Recombinant Fusion Proteins; D009964:Orotidine-5'-Phosphate Decarboxylase; D001123:Argininosuccinate Lyase; D010453:Peptide Synthases; C022379:phosphoribosylaminoimidazole-succinocarboxamide synthetase; D000432:Methanol