Case Studies - Yeast Expression
1. Protein expression in yeast
GenScript's proprietary YeastHIGHTM
expression system and in-house
YeastFast technology delivers unparalleled advantages in secretory expression. Pictured below is an enzyme expression performed using our YeastHIGHTM expression technology. After one step purification by SP-Sepharose, we find tag-free proteins at an expression level of about 20 mg/L and at purity level of over 90%. The target protein is further identified by MALDI-TOF.
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Fig. A: Expression and secretion of target protein from P.pastoris
Lane 1 : Conventinal yeast expression technology
Lane 2 : GenScript YeastHIGHTM technology
Lane 3 : Negative control
Fig. B: Purification of the target protein by SP-Sepharose
Lane M: Smart Protein Standard (GenScript, Cat. No. MM0900) |
2. Signal peptides
Shown below is the expression of a protease with signal peptide. There was no secretion expression of target protein with Saccharomyces cerevisiae α factor prepro peptide. After changing the peptide sequence for secretion from α factor prepro to the native signal peptide of the protease itself, the target protein was expressed and secreted successfully.
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Fig. A: Western blot analysis of target protein expression with native signal peptide
Lane 1-5: Induced for 24h, 48h, 72h, 84h, 120h
Fig. B: Western blot analysis of
target protein expression with α factor prepro peptide
Lane 1-4: Induced for 24h, 48h, 72h,
84h
Lane M: Protein Marker |
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