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

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, cel...

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Main Authors: Chew, Few Ne, Tan, Wen Siang, Ling, Tau Chuan, Tey, Beng Ti
Format: Article
Language:English
Published: Wiley 2009
Online Access:http://psasir.upm.edu.my/id/eprint/16730/1/Single-step%20purification%20of%20the%20recombinant%20.pdf
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spelling oai:psasir.upm.edu.my:16730 http://psasir.upm.edu.my/id/eprint/16730/ Single-step purification of the recombinant green fluorescent protein from intact Escherichia coli cells using preparative PAGE Chew, Few Ne Tan, Wen Siang Ling, Tau Chuan Tey, Beng Ti 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%. Wiley 2009-09 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/16730/1/Single-step%20purification%20of%20the%20recombinant%20.pdf 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 10.1002/elps.200900246
institution UPM IR
collection UPM IR
language English
description 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%.
format Article
author Chew, Few Ne
Tan, Wen Siang
Ling, Tau Chuan
Tey, Beng Ti
spellingShingle Chew, Few Ne
Tan, Wen Siang
Ling, Tau Chuan
Tey, Beng Ti
Single-step purification of the recombinant green fluorescent protein from intact Escherichia coli cells using preparative PAGE
author_facet Chew, Few Ne
Tan, Wen Siang
Ling, Tau Chuan
Tey, Beng Ti
author_sort Chew, Few Ne
title Single-step purification of the recombinant green fluorescent protein from intact Escherichia coli cells using preparative PAGE
title_short Single-step purification of the recombinant green fluorescent protein from intact Escherichia coli cells using preparative PAGE
title_full Single-step purification of the recombinant green fluorescent protein from intact Escherichia coli cells using preparative PAGE
title_fullStr Single-step purification of the recombinant green fluorescent protein from intact Escherichia coli cells using preparative PAGE
title_full_unstemmed Single-step purification of the recombinant green fluorescent protein from intact Escherichia coli cells using preparative PAGE
title_sort single-step purification of the recombinant green fluorescent protein from intact escherichia coli cells using preparative page
publisher Wiley
publishDate 2009
url http://psasir.upm.edu.my/id/eprint/16730/1/Single-step%20purification%20of%20the%20recombinant%20.pdf
_version_ 1819294060391694336
score 13.4562235

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