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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).
Haiyan Zhou, and
Jun Ma, and
Xueli Yang, and
Xiaohai Gong, and
Qiuping Li, and
Jian Jin
January 2017,
Methods in molecular biology (Clifton, N.J.),
Haiyan Zhou, and
Jun Ma, and
Xueli Yang, and
Xiaohai Gong, and
Qiuping Li, and
Jian Jin
June 2005,
Protein expression and purification,
Haiyan Zhou, and
Jun Ma, and
Xueli Yang, and
Xiaohai Gong, and
Qiuping Li, and
Jian Jin
July 2018,
Current protocols in molecular biology,
Haiyan Zhou, and
Jun Ma, and
Xueli Yang, and
Xiaohai Gong, and
Qiuping Li, and
Jian Jin
February 2004,
Current protocols in molecular biology,
Haiyan Zhou, and
Jun Ma, and
Xueli Yang, and
Xiaohai Gong, and
Qiuping Li, and
Jian Jin
February 2001,
Applied biochemistry and biotechnology,
Haiyan Zhou, and
Jun Ma, and
Xueli Yang, and
Xiaohai Gong, and
Qiuping Li, and
Jian Jin
August 2001,
Protein expression and purification,
Haiyan Zhou, and
Jun Ma, and
Xueli Yang, and
Xiaohai Gong, and
Qiuping Li, and
Jian Jin
January 1994,
Methods in enzymology,
Haiyan Zhou, and
Jun Ma, and
Xueli Yang, and
Xiaohai Gong, and
Qiuping Li, and
Jian Jin
March 1998,
International archives of allergy and immunology,
Haiyan Zhou, and
Jun Ma, and
Xueli Yang, and
Xiaohai Gong, and
Qiuping Li, and
Jian Jin
January 2022,
Methods in molecular biology (Clifton, N.J.),
Haiyan Zhou, and
Jun Ma, and
Xueli Yang, and
Xiaohai Gong, and
Qiuping Li, and
Jian Jin
March 2020,
Protein expression and purification,
