000 02452nam a2200265 a 4500
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005 20251117113230.0
008 141223t2014 my a f m 000 0 eng d
020 _aTHE0002251(Local)
039 9 _a201905241120
_bshamsul
_y201412231606
_zfawwaz
040 _aUMP
090 _aQP609.C39 C43 2014 r Bc.
100 3 _aChau, King Hou
245 1 0 _aElectrophoretic purification of recombinant green fluorescent protein from intact escherichia coli cells in continuous buffer system /
_cChau King Hou
260 _aKuantan, Pahang :
_bUMP,
_c2014
300 _axvi, 39 p. :
_bill. ;
_c30 cm. +
_e1 CD-ROM
500 _aFaculty of Chemical & Natural Resources Engineering
502 _aProject paper (Bachelor of Chemical Engineering) -- Universiti Malaysia Pahang – 2014
504 _aBibliography : p. 27-30
520 3 _aGreen 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
650 0 _aGreen fluorescent protein
650 0 _aEscherichia coli
999 _aVIRTUA40
_c5134
_d5140
999 _aVTLSSORT0080*0200*0400*0900*1000*2450*2600*3000*5000*5020*5040*5200*6500*6501*9992