Stability of green fluorescent protein using luminescence spectroscopy: is GFP applicable to field analysis of contaminants?
Green fluorescent protein (GFP) was first isolated in the early 1970s for experimental use from coelenterates or the Pacific jellyfish. Aequorea victoria (Morin and Hastings, 1971). GFP has since become a favored biomarker in the photophysical analysis of molecular and cell biology because of its strong intrinsic visible fluorescence and the feasibility of fusing it to other proteins without affecting their normal functions (Creemers et al., 2000). Here we report using Bacillus subtilis expressing GFP to evaluate the influence of different environmental pH conditions on GFP fluorescence. Emission acquisitions were configured to excite at 471 nm and detect at an emission from 490 to 650 nm at 1-nm increments. Fluorescence intensity was significantly better at pH 7 (4.2 x 105 cps; P-value < 0.01) than at acid or alkaline conditions. GFP is a good biomarker for environments near netural conditions: however, GFP may be unsuitable where soils or waters are below or above pH 7 because of loss in fluorescence intensity. Alternative fluorescent markers and delivery systems must be examined in different environments to optimize responses from bioreporter molecules.
Smith
C B
CB
US Army ERDC Topographic Engineering Center, Alexandria, VA 22315, USA. clint.b.smith@erdc.usace.army.mil
Anderson
J E
JE
Fischer
R L
RL
Webb
S R
SR
D001412:Bacillus subtilis / Q000737:chemistry*; D004784:Environmental Monitoring / Q000379:methods*; D004785:Environmental Pollutants / Q000032:analysis*; D049452:Green Fluorescent Proteins; D006863:Hydrogen-Ion Concentration; D008164:Luminescent Proteins / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
D004785:Environmental Pollutants; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins