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KMID : 1094720150200040800
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Biotechnology and Bioprocess Engineering
2015 Volume.20 No. 4 p.800 ~ p.806
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In Vitro evaluation and monitoring of the expression level and localization of aldose reductase using functionalized quantum dots and EGFP
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Liu Xiaomin
Yang Chengbin
Liu Jing
Liu Jianwei
Hu Rui
Lian Hongwei
Lin Guimiao
Liu Liwei
Yong Ken-Tye
Ye Ling
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Abstract
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The optimization of aldose reductase (AR) expression levels and tracking of the AR expression sites within the cell is an essential step in developing a platform for the effective production of aldose reductase inhibitors (ARIs). In this study, we have demonstrated the use of both immunocytochemistry and quantum dots-based immunofluorescence techniques for observing and detecting the expression level and localization of AR in the cytoplasm and cell membrane of a eukaryotic cell model with high levels of AR protein expression. Our results show that high expression levels of human AR can be achieved using the eukaryotic cell model that we have developed. The overexpressed AR can be used for translational studies of hAR and the screening of ARIs. More importantly, the use of the established quantum dots-based immunofluorescence technique in the intracellular labeling of AR allows the determination of the expression and distribution of the AR gene. Overall, the use of the interdisciplinary approach of both genetic engineering and quantum dot-based immunofluorescence allows not only the effective production of a desired protein, but also the determination of the cellular localization of such an expressed protein.
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KEYWORD
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aldose reductase, COS-7 cells, enhanced green fluorescence protein (EGFP), quantum dots, immunofluorescence
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