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虫号: 284986
注册: 2006-10-14
专业: 环境微生物学

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爱与雨下(金币+2): 鼓励一下！~ 2011-09-09 12:08:09
huojinlong9287(金币+60, 翻译EPI+1): 翻译的不完全 2011-09-11 09:22:48
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sltmac(金币+10): 2011-09-15 08:02:05
sltmac: 欢迎常来本版交流~ 2011-09-15 08:02:14

The aim of this study was to construct prokaryotic expression vector pET32a-AcGFP1, and its high level expression in E. coli cell. AcGFP1 gene was amplified from vector pIRES-AcGFP1 by PCR method, and was inserted into prokaryotic expression vector pET32a(+). The resulting pET32a-AcGFP1 was transformed into E. coli Rosetta(DE3) for expression followed by induction with isopropyl β-D-thiogalactopyranoside (IPTG). Whole-cell extracts were separated on 15% SDS-PAGE. AcGFP1 gene in pET32a-AcGFP1 was consistent with related gene in pIRES-AcGFP1 from Clontech Ltd, indicating prokaryotic expression vector with AcGFP1 gene was constructed successfully. High level expression was conducted with transformed E. coli Rosetta(DE3) after IPTG induction. A strong induction of a 44 kD protein was detected in SDS-PAGE, which was in correspondence with the theoretic value.

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