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Single-step purification of the recombinant green fluorescent protein from intact Escherichia coli cells using preparative PAGE


Citation

Chew, Few Ne and Tan, Wen Siang and Ling, Tau Chuan and Tey, Beng Ti (2009) Single-step purification of the recombinant green fluorescent protein from intact Escherichia coli cells using preparative PAGE. Electrophoresis, 30 (17). pp. 3017-3023. ISSN 0173-0835; ESSN: 1522-2683

Abstract / Synopsis

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%.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Engineering
DOI Number: https://doi.org/10.1002/elps.200900246
Publisher: Wiley
Keywords: Escherichia coli; Green fluorescent protein; Preparative PAGE; Protein purification
Depositing User: Nurul Ainie Mokhtar
Date Deposited: 03 Aug 2015 10:10
Last Modified: 03 Nov 2015 15:44
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1002/elps.200900246
URI: http://psasir.upm.edu.my/id/eprint/16730
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