Expression and purification of recombinant huwentoxin-I in Pichia pastoris / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 172-177, 2002.
Artigo
em Chinês
| 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.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Farmacologia
/
Pichia
/
Túbulos Seminíferos
/
Venenos de Aranha
/
Aranhas
/
Fatores de Tempo
/
Proteínas Recombinantes de Fusão
/
Dados de Sequência Molecular
/
Expressão Gênica
/
Cromatografia Líquida de Alta Pressão
Limite:
Animais
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2002
Tipo de documento:
Artigo
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