Detection of green fluorescent protein in a single bacterium by capillary electrophoresis with laser-induced fluorescence.

Full Abstract

Green fluorescence protein (GFP) is a common reporter used to monitor protein expression in single cells. However, autofluorescence from endogenous components can mask the signal from GFP, particularly at low expression levels in prokaryotes. We employ capillary electrophoresis with laser-induced fluorescence for the analysis of the expression of green fluorescent protein in a single bacterium. Capillary electrophoresis separates GFP from native cellular autofluorescent components, reducing the background signal and improving detection limits. Our system provides 100 ymol (60 copies) limits of detection for GFP. To demonstrate the performance of this instrument, we employ a model system of Deinococcus radiodurans that has been engineered to express GFP under the control of the recA promoter. We report resolution and detection of GFP and autofluorescent components in a single D. radiodurans bacterium. This paper presents the first example of expression of GFP in D. radiodurans and the first detection of GFP in a single bacterium by capillary electrophoresis.

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Author information

Author/s: Turner, Emily H (EH); Lauterbach, Kevin (K); Pugsley, Haley R (HR); Palmer, Vanessa R (VR); Dovichi, Norman J (NJ);

Affiliation: Department of Chemistry, University of Washington, Seattle, Washington 98195-1700, USA.

Journal and publication information

Publication Type: Evaluation Studies; Journal Article; Research Support, U.S. Gov't, Non-P.H.S.

Journal: Analytical chemistry (Anal Chem), published in United States. (Language: eng)

Reference: 2007-Jan; vol 79 (issue 2) : pp 778-81

Dates: Created 2007/01/15; Completed 2007/03/22;

PMID: 17222051, status: MEDLINE (last retrieval date: 12/26/2008)

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Deinococcus - chemistry; genetics
Electrophoresis, Capillary - methods
Fluorescence

Fluorescence
Green Fluorescent Proteins - analysis; genetics
Lasers

Associated Chemicals: Green Fluorescent Proteins (147336-22-9)

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