Expression and purification of hPARP1 by baculovirus system / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 998-1005, 2013.
Artigo
em Chinês
| WPRIM
| ID: wpr-233180
ABSTRACT
PARP1 is an important part of DNA repair machinery. In recent years, PARP1 as novel anti-cancer therapeutic target has been broadly explored. In this study, we expressed hPARP1 enzyme in the baculovirus system and tested its activity. We inserted hPARP1 gene into the pFastBac1, a baculovirus transfer vector and then transformed it into DH10Bac containing a shuttle vector of Bacmid. After co-transfecting the recombinant plasmid into Sf9 insect cells, the expressed hPARP1 was purified by 3-aminobezamide affinity chromatography. The expression of hPARPI was visualized by SDS-PAGE and Western blotting; the activity of expressed and purified hPARP1 was confirmed by the reaction of consumption of NAD+ by hPARP1 in vitro. After the purification by 3-aminobezamide affinity column, 3.2 mg protein was obtained and its specific activity was 1.988 nmol/(min x microg).
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Proteínas Recombinantes
/
Transfecção
/
Western Blotting
/
Baculoviridae
/
Poli(ADP-Ribose) Polimerases
/
Eletroforese em Gel de Poliacrilamida
/
Células Sf9
/
Poli(ADP-Ribose) Polimerase-1
/
Vetores Genéticos
/
Genética
Limite:
Animais
/
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2013
Tipo de documento:
Artigo
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