The analysis of heterogeneity of HWTX-I expressed in Pichia pastoris / 生物工程学报
Chinese Journal of Biotechnology
; (12): 215-219, 2006.
Artigo
em Chinês
| WPRIM (Pacífico Ocidental)
| ID: wpr-236998
Biblioteca responsável:
WPRO
ABSTRACT
To seek the reason of heterogeneity of recombinant HWTX-I (rHWTX-I) expressed in Pichia pastoris. We expressed HWTX-I gene of interest in Pichia pastoris GS115/HWTX-I. The heterogenous product expressed was separated, purified and identified by using Ion exchange HPLC, reverse HPLC, Tricine SDS-PAGE and MALDI-TOF Mass Spectrometry and then sequenced in both N-terminus and C-terminus. These results show that the heterogeneity of rHWTX-I results from the incomplete processing of signal peptide of N-terminus and the internal degradation of C-terminus. Biological activity assay shows that the activity of the heterogenous rHWTX-I only showed 30% activity compared with the native HWTX-I. The Solutions to how to avoid the heterogeneity are also discussed.
Texto completo:
Disponível
Base de dados:
WPRIM (Pacífico Ocidental)
Assunto principal:
Pichia
/
Venenos de Aranha
/
Proteínas Recombinantes
/
Proteínas de Répteis
/
Genética
/
Metabolismo
/
Neurotoxinas
Limite:
Animais
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2006
Tipo de documento:
Artigo