Expression and purification of hPARP1 by baculovirus system / 生物工程学报
Chinese Journal of Biotechnology
; (12): 998-1005, 2013.
Article
en Zh
| WPRIM
| ID: wpr-233180
Biblioteca responsable:
WPRO
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:
1
Índice:
WPRIM
Asunto principal:
Proteínas Recombinantes
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Transfección
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Western Blotting
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Baculoviridae
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Poli(ADP-Ribosa) Polimerasas
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Electroforesis en Gel de Poliacrilamida
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Células Sf9
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Poli(ADP-Ribosa) Polimerasa-1
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Vectores Genéticos
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Genética
Límite:
Animals
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Humans
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Año:
2013
Tipo del documento:
Article