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Electrophoretic purification of recombinant green fluorescent protein from intact escherichia coli cells in continuous buffer system

Green fluorescent protein (GFP) is a protein that consists of 27 kDa protein of 238 amino acid residues. GFP emits bright green fluorescence light when exposed to blue or ultraviolet light. GFP has been used as a marker for the gene expression visualization, protein localization in living and fixed...

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Main Author: Chau, King Hou
Format: Undergraduates Project Papers
Language:English
Published: 2014
Subjects:
QP Physiology
Online Access:http://umpir.ump.edu.my/id/eprint/9207/1/cd8608.pdf
http://umpir.ump.edu.my/id/eprint/9207/
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spelling my.ump.umpir.92072021-07-15T03:50:20Z http://umpir.ump.edu.my/id/eprint/9207/ Electrophoretic purification of recombinant green fluorescent protein from intact escherichia coli cells in continuous buffer system Chau, King Hou QP Physiology Green fluorescent protein (GFP) is a protein that consists of 27 kDa protein of 238 amino acid residues. GFP emits bright green fluorescence light when exposed to blue or ultraviolet light. GFP has been used as a marker for the gene expression visualization, protein localization in living and fixed tissues as well as for protein targeting in intact cells and organisms. A direct purification method was developed to purify the recombinant GFP from intact Escherichia coli (E. coli) cells using preparative native polyacrylamide gel electrophoresis (n-PAGE) in continuous buffer system. 100 μL of 12% (w/v) polyacrylamide gel was used to study the effect of biomass concentration and the effect of resolving gel height on the preparative n-PAGE. The amount of purified GFP was determined by using the gel-based imaging method and the Lowry protein determination method to determine the purity and yield of the recovered GFP. The optimal biomass concentration in the feedstock was found at 15% (w/v) with 62.5% of purity. The purity of GFP slightly reduced when the biomass concentration increased to 25% (w/v). Meanwhile, 89% of purity was achieved when 1 cm of resolving gel was employed in preparative n-PAGE. The purity of the GFP decreased when the gel height increased to 2.5cm. However, the percentage of the yield in this study was unable to determine since the calculation was completely offset 2014-01 Undergraduates Project Papers NonPeerReviewed application/pdf en http://umpir.ump.edu.my/id/eprint/9207/1/cd8608.pdf Chau, King Hou (2014) Electrophoretic purification of recombinant green fluorescent protein from intact escherichia coli cells in continuous buffer system. Faculty of Chemical & Natural Resources Engineering, Universiti Malaysia Pahang.
institution Universiti Malaysia Pahang
building UMP Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Pahang
content_source UMP Institutional Repository
url_provider http://umpir.ump.edu.my/
language English
topic QP Physiology
spellingShingle QP Physiology
Chau, King Hou
Electrophoretic purification of recombinant green fluorescent protein from intact escherichia coli cells in continuous buffer system
description Green fluorescent protein (GFP) is a protein that consists of 27 kDa protein of 238 amino acid residues. GFP emits bright green fluorescence light when exposed to blue or ultraviolet light. GFP has been used as a marker for the gene expression visualization, protein localization in living and fixed tissues as well as for protein targeting in intact cells and organisms. A direct purification method was developed to purify the recombinant GFP from intact Escherichia coli (E. coli) cells using preparative native polyacrylamide gel electrophoresis (n-PAGE) in continuous buffer system. 100 μL of 12% (w/v) polyacrylamide gel was used to study the effect of biomass concentration and the effect of resolving gel height on the preparative n-PAGE. The amount of purified GFP was determined by using the gel-based imaging method and the Lowry protein determination method to determine the purity and yield of the recovered GFP. The optimal biomass concentration in the feedstock was found at 15% (w/v) with 62.5% of purity. The purity of GFP slightly reduced when the biomass concentration increased to 25% (w/v). Meanwhile, 89% of purity was achieved when 1 cm of resolving gel was employed in preparative n-PAGE. The purity of the GFP decreased when the gel height increased to 2.5cm. However, the percentage of the yield in this study was unable to determine since the calculation was completely offset
format Undergraduates Project Papers
author Chau, King Hou
author_facet Chau, King Hou
author_sort Chau, King Hou
title Electrophoretic purification of recombinant green fluorescent protein from intact escherichia coli cells in continuous buffer system
title_short Electrophoretic purification of recombinant green fluorescent protein from intact escherichia coli cells in continuous buffer system
title_full Electrophoretic purification of recombinant green fluorescent protein from intact escherichia coli cells in continuous buffer system
title_fullStr Electrophoretic purification of recombinant green fluorescent protein from intact escherichia coli cells in continuous buffer system
title_full_unstemmed Electrophoretic purification of recombinant green fluorescent protein from intact escherichia coli cells in continuous buffer system
title_sort electrophoretic purification of recombinant green fluorescent protein from intact escherichia coli cells in continuous buffer system
publishDate 2014
url http://umpir.ump.edu.my/id/eprint/9207/1/cd8608.pdf
http://umpir.ump.edu.my/id/eprint/9207/
_version_ 1706957193738190848
score 13.211869

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