Schlechter ROJun HBernach MOso SBoyd EMuñoz-Lintz DADobson RCJRemus DMRemus-Emsermann MNP2019-01-302019-01-3020181664-302X1664-302Xhttp://hdl.handle.net/10092/16431© 2018 Schlechter, Jun, Bernach, Oso, Boyd, Muñoz-Lintz, Dobson, Remus and Remus-Emsermann. Differential fluorescent labeling of bacteria has become instrumental for many aspects of microbiological research, such as the study of biofilm formation, bacterial individuality, evolution, and bacterial behavior in complex environments. We designed a variety of plasmids, each bearing one of eight unique, constitutively expressed fluorescent protein genes in conjunction with one of four different antibiotic resistance combinations. The fluorophores mTagBFP2, mTurquoise2, sGFP2, mClover3, sYFP2, mOrange2, mScarlet-I, and mCardinal, encoding for blue, cyan, green, green–yellow, yellow, orange, red, and far-red fluorescent proteins, respectively, were combined with selectable markers conferring tetracycline, gentamicin, kanamycin, and/or chloramphenicol resistance. These constructs were cloned into three different plasmid backbones: a broad host-range plasmid, a Tn5 transposon delivery plasmid, and a Tn7 transposon delivery plasmid. The utility of the plasmids and transposons was tested in bacteria from the phyla Actinobacteria, Proteobacteria, and Bacteroidetes. We were able to tag representatives from the phylum Proteobacteria at least via our Tn5 transposon delivery system. The present study enables labeling bacteria with a set of plasmids available to the community. One potential application of fluorescently-tagged bacterial species is the study of bacteria–bacteria, bacteria–host, and bacteria–environment interactions.enCopyright © 2018 Schlechter, Jun, Bernach, Oso, Boyd, Muñoz-Lintz, Dobson, Remus and Remus-Emsermann. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.fluorophorefluorescent labelingtaggingTn5Tn7transposonJournal Article2019-01-28Fields of Research::31 - Biological sciences::3107 - Microbiology::310701 - BacteriologyFields of Research::31 - Biological sciences::3105 - Genetics::310508 - Genome structure and regulationhttps://doi.org/10.3389/fmicb.2018.03052