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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2009
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| Online Access: | http://psasir.upm.edu.my/id/eprint/16730/1/Single-step%20purification%20of%20the%20recombinant%20.pdf |
| _version_ | 1796969113061425152 |
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| author | Chew, Few Ne Tan, Wen Siang Ling, Tau Chuan Tey, Beng Ti |
| author_facet | Chew, Few Ne Tan, Wen Siang Ling, Tau Chuan Tey, Beng Ti |
| author_sort | Chew, Few Ne |
| collection | UPM |
| 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%. |
| first_indexed | 2024-03-06T07:38:17Z |
| format | Article |
| id | upm.eprints-16730 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T07:38:17Z |
| publishDate | 2009 |
| publisher | Wiley |
| record_format | dspace |
| spelling | upm.eprints-167302015-11-03T07:44:08Z 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 |
| 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 |
| title | 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_short | 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 |
| url | http://psasir.upm.edu.my/id/eprint/16730/1/Single-step%20purification%20of%20the%20recombinant%20.pdf |
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