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Gateway RFP-fusion vectors for high-throughput functional analysis of genes

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Authors
Park, Jae-Yong; Hwang, Eun Mi; Park, Nammi; Kim, Eunju; Kim, Dong-Gyu; Kang, Dawon; Han, Jaehee; Choi, Wan Sung; Ryu, Pan Dong; Hong, Seong-Geun
Issue Date
2007
Publisher
한국분자·세포생물학회 = Korean Society of Molecular and Cellular Biology
Citation
Mol. Cells 23, 357-362
Keywords
Gateway Cloning SystemRed Fluorescent ProteinHigh Throughput Screening System
Abstract
There is an increasing demand for high throughput
(HTP) methods for gene analysis on a genome-wide
scale. However, the current repertoire of HTP detection
methodologies allows only a limited range of cellular
phenotypes to be studied. We have constructed two
HTP-optimized expression vectors generated from the
red fluorescent reporter protein (RFP) gene. These vectors
produce RFP-tagged target proteins in a multiple
expression system using gateway cloning technology
(GCT). The RFP tag was fused with the cloned genes,
thereby allowing us localize the expressed proteins in
mammalian cells. The effectiveness of the vectors was
evaluated using an HTP-screening system. Sixty representative
human C2 domains were tagged with RFP and
overexpressed in HiB5 neuronal progenitor cells, and
we studied in detail two C2 domains that promoted the
neuronal differentiation of HiB5 cells. Our results show
that the two vectors developed in this study are useful
for functional gene analysis using an HTP-screening
system on a genome-wide scale.
ISSN
1016-8478
Language
English
URI
http://molcells.inforang.com/about/journal_view.html?uid=257

http://hdl.handle.net/10371/7031
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College of Veterinary Medicine (수의과대학)Dept. of Veterinary Medicine (수의학과)Journal Papers (저널논문_수의학과)
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