Abstract
The utility of the green fluorescent protein (GFP) as a probe to monitor protein localization in living cells is gaining a great deal of attention. In this study, to understand the localization of luciferases in E. coli, we have attached GFP tags at both the N- and the C-terminus of firefly luciferase (FF-Luc) (from Pyrocoelia miyako) and of red (RE-Luc) and green (GR-Luc) bioluminescence-emitting luciferases (from Phrixothrix railroad-worms), respectively. There was no significant change in the bioluminescence emission spectrum for any of the three luciferases following the tagging with GFP at either the N- or C-terminus, confirming the absence of energy transfer between one another. Using confocal imaging microscopy, we observed that all three luciferases expressed in the E.coli cultured at 37 °C tend to aggregate and are seen to localize in the poles, thus confirming their poor folding properties. In contrast, in the E.coli cultured at 18 °C FF-Luc was found to be highly expressed in the soluble form when compared to RE-Luc and GR-Luc. These results support our previous finding that the folding properties of FF-Luc and RE/GR-Luc are totally different.
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Presented at the 14th International Congress on Photobiology, at Jungmoon, Jeju Island, South Korea, 10th-15th June 2004.
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Venkatesh, B., Arifuzzaman, M., Mori, H. et al. Use of GFP tags to monitor localization of different luciferases in E. coli. Photochem Photobiol Sci 4, 740–743 (2005). https://doi.org/10.1039/b416747c
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DOI: https://doi.org/10.1039/b416747c