Hormone-inducible expression and metal affinity chromatography of recombinant proteins in Saccharomyces cerevisiae.
An economical method to express recombinant protein in yeast (Saccharomyces cerevisiae) was developed. We combined two principles: Ni(2+)-agarose-based affinity purification of fusion proteins and expression from a cassette regulated by steroid hormones. After induction by desoxycorticosterone, > 90% pure protein was obtained after a single round of metal affinity chromatography. Gentle lysis conditions allowed the preservation of enzymatic activity. Average yields of 10 mg protein per liter of culture were obtained at a fraction of the cost of other eukaryotic expression systems.
Clausen
M
M
Plant Biology Lab, Salk Institute, San Diego, California 92186-5800.
Lamb
C J
CJ
Megnet
R
R
Doerner
P W
PW
D001483:Base Sequence; D002262:Carboxy-Lyases / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000235:genetics / Q000302:isolation & purification*; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000235:genetics; D005822:Genetic Vectors; D006728:Hormones / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000187:drug effects; D009532:Nickel; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012685:Sepharose
D005656:Fungal Proteins; D006728:Hormones; D012333:RNA, Messenger; D011994:Recombinant Proteins; D009532:Nickel; D012685:Sepharose; D002262:Carboxy-Lyases