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KMID : 0923620090090060243
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Immune Network
2009 Volume.9 No. 6 p.243 ~ p.247
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A New Reporter Vector System Based on Flow-Cytometry to Detect Promoter Activity
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Jung Sun-Do
Choi Ji-Hye
Hong Chang-Wan
Lee Hyun-Ji
Park Yoon-Kyung
Shin Jung-Hoon
Park Se-Ho
Park Jae-Won
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Abstract
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In this study, we report the development of a new dual reporter vector system for the analysis of promoter activity. This system employs green fluorescence emitting protein, EGFP, as a reporter, and uses red fluorescence emitting protein, DsRed, as a transfection control in a single vector. The expression of those two proteins can be readily detected via flow cytometry in a single analysis, with no need for any further manipulation after transfection. As this system allows for the simultaneous detection of both the control and reporter proteins in the same cells, only transfected cells which express the control protein, DsRed, can be subjected to promoter activity analysis, via the gating out of all un-transfected cells. This results in a dramatic increase in the promoter activity detection sensitivity. This novel reporter vector system should prove to be a simple and efficient method for the analysis of promoter activity.
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KEYWORD
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Dual reporter vector system, Transcription activity, Promoter assay, FACS
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