快速纯化在大肠杆菌中表达的增强型绿色荧光蛋白

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快速纯化在大肠杆菌中表达的增强型绿色荧光蛋白

周笑鹏^a; 史清洪^a; 邢新会^b; 孙彦^a

^a Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China
^b Department of Chemical Engineering, Tsinghua University, Beijing 100084, China

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2006-04-28 发布日期:2006-04-28
通讯作者: 孙彦

Rapid Purification of Enhanced Green Fluorescent Protein from
Escherichia coli

ZHOU Xiaopeng^a; SHI Qinghong^a; XING Xinhui^b; SUN Yan^a

^a Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China
^b Department of Chemical Engineering, Tsinghua University, Beijing 100084, China

Received:1900-01-01 Revised:1900-01-01 Online:2006-04-28 Published:2006-04-28
Contact: SUN Yan

摘要/Abstract

摘要： As an excellent reporter molecule, enhanced green fluorescent protein (eGFP) was widely used for gene expression and regulation and was generally expressed in Escherichia coli strain. A rapid procedure consisting of ammonium sulfate precipitation, size exclusion chromatography, and anion exchange chromatography was developed for the purification of eGFP. Based on the proposed procedure, recombinant eGFP with an electrophoretic purity was achieved in combination with an overall yield of 66% and a purification factor of 17.9. The fluorescent spectrometry of purified eGFP and lysate from E. coli strain expressing eGFP exhibited the same wavelength of ex-citation and emission maxima, indicating that the purification procedure did not influence the construct and fluo-rescent characteristics of desired protein. The procedure mentioned was easy to scale up for the purification of large quantities of eGFP

关键词: enhanced green fluorescent protein;purification;size exclusion chromatography;anion exchange chromatography

Abstract: As an excellent reporter molecule, enhanced green fluorescent protein (eGFP) was widely used for gene expression and regulation and was generally expressed in Escherichia coli strain. A rapid procedure consisting of ammonium sulfate precipitation, size exclusion chromatography, and anion exchange chromatography was developed for the purification of eGFP. Based on the proposed procedure, recombinant eGFP with an electrophoretic purity was achieved in combination with an overall yield of 66% and a purification factor of 17.9. The fluorescent spectrometry of purified eGFP and lysate from E. coli strain expressing eGFP exhibited the same wavelength of ex-citation and emission maxima, indicating that the purification procedure did not influence the construct and fluo-rescent characteristics of desired protein. The procedure mentioned was easy to scale up for the purification of large quantities of eGFP

Key words: enhanced green fluorescent protein, purification, size exclusion chromatography, anion exchange chromatography

周笑鹏, 史清洪, 邢新会, 孙彦. 快速纯化在大肠杆菌中表达的增强型绿色荧光蛋白
[J]. , DOI: .

ZHOU Xiaopeng, SHI Qinghong, XING Xinhui, SUN Yan. Rapid Purification of Enhanced Green Fluorescent Protein from
Escherichia coli
[J]. , DOI: .

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