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

Jae-Yong Park, Eun Mi Hwang, Nammi Park, Eunju Kim, Dong Gyu Kim, Dawon Kang, Jaehee Han, Wan Sung Choi, Pan Dong Ryu, Seong Geun Hong

Research output: Contribution to journalArticle

7 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)357-362
Number of pages6
JournalMolecules and cells
Volume23
Issue number3
Publication statusPublished - 2007 Jun 30
Externally publishedYes

Keywords

  • Gateway cloning system
  • High throughput screening system
  • Red fluorescent protein

ASJC Scopus subject areas

  • Medicine(all)

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  • Cite this

    Park, J-Y., Hwang, E. M., Park, N., Kim, E., Kim, D. G., Kang, D., Han, J., Choi, W. S., Ryu, P. D., & Hong, S. G. (2007). Gateway RFP-fusion vectors for high throughput functional analysis of genes. Molecules and cells, 23(3), 357-362.