Production of mitotic regulators using an autoselection system for protein expression in budding yeast.
Methods Mol Biol
; 545: 63-80, 2009.
Article
in En
| MEDLINE
| ID: mdl-19475382
ABSTRACT
A novel protein expression system in budding yeast is described which has been used to express many yeast mitotic regulators as well as a wide range of other recombinant proteins from several different species. The expression system relies on autoselection with essential genes to maintain high copy numbers of expression plasmids. Autoselection permits expression cells to be grown in rich medium with no need for plasmid selection with drugs or nutritional conditions. This optimizes growth and expression of recombinant proteins. The use of the expression system is illustrated by purifying budding yeast mitotic regulators, Cdc14 and Net1, and recapitulating their activities in vitro.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Saccharomyces cerevisiae
/
Protein Tyrosine Phosphatases
/
Cell Cycle Proteins
/
Saccharomyces cerevisiae Proteins
Language:
En
Journal:
Methods Mol Biol
Journal subject:
BIOLOGIA MOLECULAR
Year:
2009
Document type:
Article
Affiliation country:
United kingdom