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Journal of Applied Sciences
  Year: 2014 | Volume: 14 | Issue: 5 | Page No.: 489-492
DOI: 10.3923/jas.2014.489.492
Identification of Transfection Efficiency Using Qualitative and Quantitative Analyses of Green Fluorescent Protein in CHO Cells
A. Nurul Atikah, Z. Zulkiflie, M.A.W. Rohaya, S. Sahidan, Z.A. Intan Zarina, M.Y. Nurul Yuziana, M. Zulkifli and Z.A. Shahrul Hisham

Abstract:
Tranfection is a method that randomly introduce foreign DNA into cells either to produce genetically modified cells or to produce recombinant protein. However, efficient transfection is needed for expression of recombinant protein in mammalian cell. This study aim is to identify the transfection efficiency of novel expression vectors by qualitative and quantitative analyses. The novel expression vectors were named, pZAAGFP (without interested element), pZAAH1C (with integrated element) and pZAAM956 (with enhancer element). Chinese Hamster Ovary (CHO) cells were transfected with 3 different novel expression vectors by using Lipofectamine LTX. After 24 h, these cells were observed under confocal fluorescence microscope. The frequency of both transfected and non-tranfected cells were determined. Then, the intensity of Green Fluorescent Protein (GFP) was quantified using fluorescence reader at emission wavelength 506 nm and excitation wavelength 500 nm. Qualitative analysis showed that all novel expression vectors were able to express GFP. More than 80% CHO cells had been successfully transfected with all of the novel expression vectors stated. Quantitative analysis also showed that GFP intensity of both novel expression vectors named, pZAAH1C and pZAAM956 were higher than commercial vector and the GFP intensity of pZAAGFP was about the same as commercial vector. This finding indicates that all novel expression vectors are able to express the GFP gene and both qualitative and quantitative analyses can be applied to determine transfection efficiency.
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How to cite this article:

A. Nurul Atikah, Z. Zulkiflie, M.A.W. Rohaya, S. Sahidan, Z.A. Intan Zarina, M.Y. Nurul Yuziana, M. Zulkifli and Z.A. Shahrul Hisham, 2014. Identification of Transfection Efficiency Using Qualitative and Quantitative Analyses of Green Fluorescent Protein in CHO Cells. Journal of Applied Sciences, 14: 489-492.

DOI: 10.3923/jas.2014.489.492

URL: https://scialert.net/abstract/?doi=jas.2014.489.492

 
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