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Single‐step purification of the recombinant green fluorescent protein from intact Escherichia coli cells using preparative PAGE
Author(s) -
Chew Few Ne,
Tan Wen Siang,
Ling Tau Chuan,
Tey Beng Ti
Publication year - 2009
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200900246
Subject(s) - green fluorescent protein , escherichia coli , recombinant dna , intracellular , fluorescence , polyacrylamide gel electrophoresis , yield (engineering) , chemistry , chromatography , protein purification , biochemistry , bacteria , cell disruption , microbiology and biotechnology , biology , gene , enzyme , materials science , physics , genetics , quantum mechanics , metallurgy
Mechanical and non‐mechanical breakages of bacterial cells are usually the preliminary steps in intracellular protein purification. In this study, the recombinant green fluorescent protein (GFP) was purified from intact Escherichia coli cells using preparative PAGE. In this purification process, cells disruption step is not needed. The cellular content of E. coli was drifted out electrically from cells and the negatively charged GFP was further electroeluted from polyacrylamide gel column. SEM investigation of the electrophoresed cells revealed substantial structural damage at the cellular level. This integrated purification technique has successfully recovered the intracellular GFP with a yield of 82% and purity of 95%.