Cargando…

Imaging living obligate anaerobic bacteria with bilin-binding fluorescent proteins

Fluorescent tools such as green fluorescent protein (GFP) have been used extensively as reporters in biochemistry and microbiology, but GFP and other conventional fluorescent proteins are restricted to aerobic environments. This limitation precludes fluorescence studies of anaerobic ecologies includ...

Descripción completa

Detalles Bibliográficos
Autores principales: Chia, Hannah E., Zuo, Tiancheng, Koropatkin, Nicole M., Marsh, E. Neil G., Biteen, Julie S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7731933/
https://www.ncbi.nlm.nih.gov/pubmed/33313576
http://dx.doi.org/10.1016/j.crmicr.2020.04.001
_version_ 1783621996162056192
author Chia, Hannah E.
Zuo, Tiancheng
Koropatkin, Nicole M.
Marsh, E. Neil G.
Biteen, Julie S.
author_facet Chia, Hannah E.
Zuo, Tiancheng
Koropatkin, Nicole M.
Marsh, E. Neil G.
Biteen, Julie S.
author_sort Chia, Hannah E.
collection PubMed
description Fluorescent tools such as green fluorescent protein (GFP) have been used extensively as reporters in biochemistry and microbiology, but GFP and other conventional fluorescent proteins are restricted to aerobic environments. This limitation precludes fluorescence studies of anaerobic ecologies including polymicrobial communities in the human gut microbiome and in soil microbiomes, which profoundly affect health, disease, and the environment. To address this limitation, we describe the first implementation of two bilin-binding fluorescent proteins (BBFPs), UnaG and IFP2.0, as oxygen-independent fluorescent labels for live-cell imaging in anaerobic bacteria. Expression of UnaG or IFP2.0 in the prevalent gut bacterium Bacteroides thetaiotaomicron (B. theta) results in detectable fluorescence upon the addition of the bilirubin or biliverdin ligand, even in anaerobic conditions. Furthermore, these BBFPs can be used in two-color imaging to differentiate cells expressing either UnaG or IFP2.0; UnaG and IFP2.0 can also be used to distinguish B. theta from other common gut bacterial species in mixed-culture live-cell imaging. BBFPs are promising fluorescent tools for live-cell imaging investigations of otherwise inaccessible anaerobic polymicrobial communities.
format Online
Article
Text
id pubmed-7731933
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-77319332020-12-11 Imaging living obligate anaerobic bacteria with bilin-binding fluorescent proteins Chia, Hannah E. Zuo, Tiancheng Koropatkin, Nicole M. Marsh, E. Neil G. Biteen, Julie S. Curr Res Microb Sci Research Paper Fluorescent tools such as green fluorescent protein (GFP) have been used extensively as reporters in biochemistry and microbiology, but GFP and other conventional fluorescent proteins are restricted to aerobic environments. This limitation precludes fluorescence studies of anaerobic ecologies including polymicrobial communities in the human gut microbiome and in soil microbiomes, which profoundly affect health, disease, and the environment. To address this limitation, we describe the first implementation of two bilin-binding fluorescent proteins (BBFPs), UnaG and IFP2.0, as oxygen-independent fluorescent labels for live-cell imaging in anaerobic bacteria. Expression of UnaG or IFP2.0 in the prevalent gut bacterium Bacteroides thetaiotaomicron (B. theta) results in detectable fluorescence upon the addition of the bilirubin or biliverdin ligand, even in anaerobic conditions. Furthermore, these BBFPs can be used in two-color imaging to differentiate cells expressing either UnaG or IFP2.0; UnaG and IFP2.0 can also be used to distinguish B. theta from other common gut bacterial species in mixed-culture live-cell imaging. BBFPs are promising fluorescent tools for live-cell imaging investigations of otherwise inaccessible anaerobic polymicrobial communities. Elsevier 2020-05-01 /pmc/articles/PMC7731933/ /pubmed/33313576 http://dx.doi.org/10.1016/j.crmicr.2020.04.001 Text en © 2020 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Paper
Chia, Hannah E.
Zuo, Tiancheng
Koropatkin, Nicole M.
Marsh, E. Neil G.
Biteen, Julie S.
Imaging living obligate anaerobic bacteria with bilin-binding fluorescent proteins
title Imaging living obligate anaerobic bacteria with bilin-binding fluorescent proteins
title_full Imaging living obligate anaerobic bacteria with bilin-binding fluorescent proteins
title_fullStr Imaging living obligate anaerobic bacteria with bilin-binding fluorescent proteins
title_full_unstemmed Imaging living obligate anaerobic bacteria with bilin-binding fluorescent proteins
title_short Imaging living obligate anaerobic bacteria with bilin-binding fluorescent proteins
title_sort imaging living obligate anaerobic bacteria with bilin-binding fluorescent proteins
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7731933/
https://www.ncbi.nlm.nih.gov/pubmed/33313576
http://dx.doi.org/10.1016/j.crmicr.2020.04.001
work_keys_str_mv AT chiahannahe imaginglivingobligateanaerobicbacteriawithbilinbindingfluorescentproteins
AT zuotiancheng imaginglivingobligateanaerobicbacteriawithbilinbindingfluorescentproteins
AT koropatkinnicolem imaginglivingobligateanaerobicbacteriawithbilinbindingfluorescentproteins
AT marsheneilg imaginglivingobligateanaerobicbacteriawithbilinbindingfluorescentproteins
AT biteenjulies imaginglivingobligateanaerobicbacteriawithbilinbindingfluorescentproteins