军事医学2016，Vol.40Issue(10)：795-800,818,7.DOI:10.7644/j.issn.1674-9960.2016.10.005
串联杂种泛素结合结构域蛋白（ThUBD）在大肠杆菌中的可溶性高效表达
Soluble expression of tandem hybrid ubiquitin-binding domains (ThUBD) in prokaryotic cytoplasm of Escherichia coli BL21(DE3)
摘要
Abstract
Objective We increase the soluble expression of artificial tandem hybrid ubiquitin binding domains ( ThUBD) in prokaryotic cytoplasm of Escherichia coli BL21 ( DE3 ) , which offer an effective and special profiling for ubiquitin conjugates( UbC) .Methods Codon optimization of the ThUBD was performed, followed by analysis of codon relative adaptiveness based on relative frequency of synonymous codon ( RFSC) of E.coli.Further induced expression and yeast ubiquitin conjugate enrichment quantified the soluble ThUBD-S and tested the ability to bind UbC.Results The statistical result showed that the percentage of codon of the highest usage frequency was increased from 48%to 75%, and codon adaptation index( CAI) was increased from 0.63 to 0.88 after codon optimization, which might suggest a higher expression of the ThUBD in E.coli BL21 (DE3).The subsequent SDS-PAGE indicated that the soluble target protein was increased four times, which accounted for 13.06%of total cell lysis.Further ubiquitinated proteome of yeast demonstrated that the ability to bind and enrich UbC of optimized ThUBD-S did not change compared with original ThUBD.Conclusion The expression of ThUBD-S can quadruple after codon optimization.At the same time, codon optimization does not impact its soluble expression and the ability to bind UbC.关键词
ThUBD/泛素化蛋白/大肠杆菌/密码子优化/融合表达/蛋白质组学Key words
ThUBD/ubiquitin conjugates(UbC)/Escherichia coli/codon optimization/recombinant expression/proteomics分类
生物科学引用本文复制引用
罗维佳,邓晨,李衍常,高媛,徐平..串联杂种泛素结合结构域蛋白（ThUBD）在大肠杆菌中的可溶性高效表达[J].军事医学,2016,40(10):795-800,818,7.基金项目
国家重点研发计划资助项目（2016YFA0501300）；国家自然科学基金资助项目（31670834）；国家863计划资助项目（2014AA020900，2014AA020607）；北京市自然科学基金资助项目（5152008）；北京百名科技领军人才计划资助项目 （）
