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Development of the yeast Pichia pastoris as a model organism for a genetic and molecular analysis of peroxisome assembly
Author(s) -
Gould Stephen J.,
McCollum Dannel,
Spong Allan P.,
Heyman John A.,
Subramani Suresh
Publication year - 1992
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320080805
Subject(s) - biology , pichia pastoris , yeast , model organism , organism , peroxisome , computational biology , genetics , pichia , evolutionary biology , gene , recombinant dna
We describe the isolation of mutants of the yeast Pichia pastoris that are deficient in peroxisome assembly ( pas ). These mutants of P. pastoris can be identified solely by their inability to grow on methanol and oleic acid, the utilization of which requires peroxisomal enzymes, and are defined by the absence of normal peroxisomes as judged by electron microscopy and biochemical fractionation experiments. These mutants are the result of genetic defects at single loci and represent at least eight different complementation groups. The isolation of pas mutants of P. pastoris by a simple screen for mutants unable to use methanol and oleic acid represents a significantly more efficient method for identification of pas mutants than is possible in other organisms. To exploit this advantage fully we also developed new reagents for the genetic and molecular manipulation of P. pastoris . These include a set of auxotropic strains with an essentialiy wild type genetic background, plasmids that act as Escherichia coli–P. pastoris shuttle vectors, and genomic DNA libraries for isolation of P. pastoris genes by functional complementation of mutants or by nucleic acid hybridization. The availability of numerous pas mutants and the reagents necessary for their molecular analysis should lead to the isolation and characterization of genes involved in peroxisome assembly.