Expression and purification of recombinant huwentoxin-I in Pichia pastoris / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 172-177, 2002.
Article
in Chinese
| WPRIM
| ID: wpr-231356
ABSTRACT
HWTX-I is a peptide neurotoxin purified from the crude venom of the Chinese bird Spider Selenocosmia Huwena, which has analyesic activity. rHWTX-I expressed by P. pastoris and secreted to culture supernatant was first precipitated by (NH4)2SO4, then it was isolated and desalted by ultrofiltration following by ion exchange chromatography of CM column, after reverse phase HPLC of C18 column and vacuum drying, the pure HWTX-I protein was obtained which was proved to be recombinant HWTX-I by Tricine SDS-PAGE, MALDI-TOF mass spectrometry, amino acid composition analysis, the N-terminal amino acid sequence and its biological activity. The final yield of the purified HWTX-I was about 80 mg/L accounting for 23.6% of its total secretory proteins.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Pichia
/
Seminiferous Tubules
/
Spider Venoms
/
Spiders
/
Time Factors
/
Recombinant Fusion Proteins
/
Molecular Sequence Data
/
Gene Expression
/
Chromatography, High Pressure Liquid
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2002
Type:
Article
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