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Proceedings Paper

Spectroscopy detection of green and red fluorescent proteins in genetically modified plants using a fiber optics system
Author(s): Oi Wah Liew; Anand Krishna Asundi; Jun-Wei Chen; Yiwen Chew; Shangjuan Yu; Gare Hoon Yeo
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Paper Abstract

In this paper, fiber optic spectroscopy is developed to detect and quantify recombinant green (EGFP) and red (DsRED) fluorescent proteins in vitro and in vivo. The bacterial expression vectors carrying the coding regions of EGFP and DsRED were introduced into Escherichia coli host cells and fluorescent proteins were produced following induction with IPTG. Soluble EGFP and DsRED proteins were isolated from lysed bacterial cells and serially diluted for quantitative analysis by fiber optic spectroscopy. Fluorescence at the appropriate emission wavelengths could be detected up to 64X dilution for EGFP and 40X dilution for DsRED. To determine the capability of spectroscopy detection in vivo, transgenic potato hairy roots expressing EGFP and DsRED were regenerated. This was achieved by cloning the EGFP and DsRED genes into the plant binary vector, pTMV35S, to create the recombinant vectors pGLOWGreen and pGLOWRed. These latter binary vectors were introduced into Agrobacterium rhizogenes strain A4T. Infection of potato cells with transformed agrobacteria was used to insert the fluorescent protein genes into the potato genome. Genetically modified potato cells were then regenerated into hairy roots. A panel of transformed hairy roots expressing varying levels of fluorescent proteins was selected by fluorescence microscopy. We are now assessing the capability of spectroscopic detection system for in vivo quantification of green and red fluorescence levels in transformed roots.

Paper Details

Date Published: 8 May 2001
PDF: 4 pages
Proc. SPIE 4416, Optical Engineering for Sensing and Nanotechnology (ICOSN 2001), (8 May 2001); doi: 10.1117/12.427067
Show Author Affiliations
Oi Wah Liew, Singapore Polytechnic (Singapore)
Anand Krishna Asundi, Nanyang Technological Univ. (Singapore)
Jun-Wei Chen, Nanyang Technological Univ. (Singapore)
Yiwen Chew, Singapore Polytechnic (Singapore)
Shangjuan Yu, Singapore Polytechnic (Singapore)
Gare Hoon Yeo, Singapore Polytechnic (Singapore)

Published in SPIE Proceedings Vol. 4416:
Optical Engineering for Sensing and Nanotechnology (ICOSN 2001)
Koichi Iwata, Editor(s)

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