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Prokaryotic Expression and Protein Purification of GmERF6 Gene(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2011年06期
Page:
906-909
Research Field:
Publishing date:

Info

Title:
Prokaryotic Expression and Protein Purification of GmERF6 Gene
Author(s):
ZHAI YingLEI Ting-tingYAN FanLI Yan-jieLI Xiao-weiWANG YingLI Jing-wenWANG Qing-yu
College of Plant Science,Jilin University,Changchun 130062,Jilin,China
Keywords:
SoybeanGmERF6Construction of vectorProkaryotic expressionPurification
PACS:
S565.1
DOI:
10.11861/j.issn.1000-9841.2011.06.0906
Abstract:
Ethylene-responsive factors(ERFs)belong to a large family of transcription factors that are specific to plants,ERFs are extensively involved in the responses to biotic and abiotic stresses.A new ERF transcription factor-GmERF6 in soybean was subcloned into prokaryotic expression vector pET28 to get the expression protein for further study.The constructed vector pET28-GmERF6 was transformed into Rosetta(DE3)to get the recombinant protein by IPTG induction.The result indicated that an 30 kDa recombinant protein was expressed when the concentration and induction time of IPTG were 0.3 mmol·L-1and 3 h,respectively,the recombinant protein was mainly existed in inclusion body form through SDS-PAGE analysis.After the inclusion body was dissolved with 8 mol·L-1 urea,the high quality recombinant protein was obtained through Ni column purification and its concentration was 0.56 mg·mL-1 after determined by Bradford method.

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Last Update: 2014-08-16