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《Acta Agriculturae Zhejiangensis》 2014-01
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Expression and purification of the novel Resilin-R5 fusion protein in Escherichia coli

MAO Zheng-zheng;LIU Yong;SUN Yan;ZHOU Man;LIU Zi-duo;TANG Jiang-wu;State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University;Institute of Plant Protection and Microbiology,Zhejiang Academy of Agricultural Sciences;Hangzhou Normal University Qianjiang College;College of Pharmacy,Zhejiang University of Technology;  
A fusion protein Resilin-R5 which was expected to have a superiority of high elasticity and silicon deposition was designed and expressed in Escherichia coli. At the same time,a facile purification method was built to obtain the new fusion protein. Firstly,the recombinant resilin-r5 gene,which constructed by fusing the diatoms R5 peptide gene sequences to the 3'end of Drosophila melanogaster resilin gene sequences,was synthesized after a rare codon optimization. Then,the new gene was inserted into the prokaryotic expression vector pET28a. The expression vector was transformed into E. coli BL21( DE3) pLysS strain,and the transformant was cultured in an auto-induced medium to get resilin-r5 gene expressed. At last,the His-tagged recombinant fusion protein was purified by‘saltingout and heating'method and Ni ion column affinity chromatography. The efficient expression of the fusion Resilin-R5 reached 120 mg per liter culture. A solid foundation was built for characterization and innovative applications of the fusion protein Resilin-R5 which has high elasticity and silicon deposition ability.
【Fund】: 浙江省农业科学院科技创新能力提升工程项目“新型载药融合蛋白制备及药物缓释性能研究”;; 浙江省自然科学基金项目“基于新型高弹蛋白-R5肽融合蛋白-SiO2复合纳米材料的控释给药系统的构建研究(LY12E03009)”;; 杭州师范大学钱江学院科学研究基金“抗菌肽-SiO2纳米复合材料的制备研究(2013QJJL05)”
【CateGory Index】: Q78
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1 Hu Renjian;Cai Jiali;Liu Li;Tu Manyu;Xu Tao;Du Cuirong;Luo Jia;Ding Sen;School of Pharmacy and Bioengineering,Chongqing University of Technology;School of Life Science,Nanjing University;;Construction of HPV18E7 recombinant plasmid and exploration of its optimization expression condition in Escherichia coli[J];Chongqing Medicine;2013-30
2 XIA Lan;Zhejiang Provincial Key Laboratory of Bio-metrology,Inspection and Quarantine,College of Life Science,China Jiliang University;;Application of Penicillin-binding Proteins in Residues Detection[J];Journal of Anhui Agricultural Sciences;2013-19
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4 LIU Xiang;LI Hui;School of Life Sciences,Sun Yat-sen University;College of Biological Sciences and Engineering,Shanxi University of Technology;;Construction of E.coli Outer Protein OmpA Expression Vector and Optimization of Inducing Conditions[J];Biotechnology;2013-04
5 ZHOU Chuanwen;LI Yipeng;DING Yu;School of Life Sciences,Fudan University;;Expression,Purification and Characterization of Recombinant Human Heme Oxygenase 2 in Escherichia coli[J];Journal of Yunnan Agricultural University(Natural Science);2013-06
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