Abstract
Objectives
To determine the effectiveness of evoglow-Pp1 as a reporter to study gene expression in bifidobacteria. To choose a strong and constitutive promoter to track fluorescently labelled bifidobacteria in environments under anaerobic conditions.
Results
The elongation factor P (EF-P) promoter from Bifidobacterium longum CECT 4551 produced the highest emission of fluorescence signal and was therefore able to produce the highest gene expression of the promoters studied. The promoters from B. longum CECT 4551 showed different fluorescence signal intensities which, in descending order, were: EF-P, initiation factor IF-2, elongation factor G, elongation factor Tu, elongation factor Nus A, elongation factor Ts and 30S ribosomal protein S12.
Conclusions
The consistency of the methods employed (fluorescence imaging system, fluorescence microscopy, fluorimetry and flow cytometry) showed that the construction pNZ:Prom.GFPana contained the anaerobic fluorescent protein evoglow-Pp1 could be exploited as a tool for analysing the gene expression in bifidobacteria strains.
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Acknowledgments
This work was supported by projects RTA2010-00116-00-00 and RM2012-00004-00-00. J. M. Landete has a postdoctoral contract with the research program “Ramón y Cajal” (MINECO, Spain).
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Montenegro-Rodríguez, C., Peirotén, A., Sanchez-Jimenez, A. et al. Analysis of gene expression of bifidobacteria using as the reporter an anaerobic fluorescent protein. Biotechnol Lett 37, 1405–1413 (2015). https://doi.org/10.1007/s10529-015-1802-8
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DOI: https://doi.org/10.1007/s10529-015-1802-8