Cloning and characterization of the Pichia pastoris MET2 gene as a selectable marker.
We describe the isolation and characterization of a new biosynthetic gene, MET2, from the methylotrophic yeast Pichia pastoris. The predicted product of PpMET2 is significantly similar to its Saccharomyces cerevisiae counterpart, ScMET2, which encodes homoserine-O-transacetylase. The ScMET2 was able to complement the P. pastoris met2 strain; however, the converse was not true. Expression vectors based on PpMET2 for the intracellular and secreted production of foreign proteins and corresponding auxotrophic strains were constructed and tested for use in heterologous expression. The expression vectors and corresponding strains provide greater flexibility when using P. pastoris for recombinant protein expression.
Thor
Der
D
Department of Biological Sciences, University of the Pacific, 3601 Pacific Avenue, Stockton, CA 95211, USA.
Xiong
See
S
Orazem
Claire C
CC
Kwan
An-Chun
AC
Cregg
James M
JM
Lin-Cereghino
Joan
J
Lin-Cereghino
Geoff P
GP
D000123:Acetyltransferases / Q000096:biosynthesis / Q000235:genetics*; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D005800:Genes, Fungal; D005819:Genetic Markers; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D010957:Plasmids; D016415:Sequence Alignment
D005819:Genetic Markers; D000123:Acetyltransferases; C021369:homoserine O-acetyltransferase