Open AccessCloning and characterization of the Pichia pastoris MET2 gene as a selectable marker
Author(s) -
Thor Der,
Xiong See,
Orazem Claire C.,
Kwan AnChun,
Cregg James M.,
LinCereghino Joan,
LinCereghino Geoff P.
Publication year - 2005
Publication title -
fems yeast research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 92
eISSN - 1567-1364
pISSN - 1567-1356
DOI - 10.1016/j.femsyr.2005.03.009
Subject(s) - pichia pastoris , biology , selectable marker , heterologous expression , auxotrophy , cloning (programming) , recombinant dna , gene , pichia , yeast , genetics , microbiology and biotechnology , plasmid , escherichia coli , computer science , programming language
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.