Cargando…
Monitoring Gene Expression during a Galleria mellonella Bacterial Infection
Galleria mellonella larvae are an alternative in vivo model that has been extensively used to study the virulence and pathogenicity of different bacteria due to its practicality and lack of ethical constraints. However, the larvae possess intrinsic autofluorescence that obstructs the use of fluoresc...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7697238/ https://www.ncbi.nlm.nih.gov/pubmed/33207842 http://dx.doi.org/10.3390/microorganisms8111798 |
_version_ | 1783615568865132544 |
---|---|
author | Moya-Andérico, Laura Admella, Joana Fernandes, Rodrigo Torrents, Eduard |
author_facet | Moya-Andérico, Laura Admella, Joana Fernandes, Rodrigo Torrents, Eduard |
author_sort | Moya-Andérico, Laura |
collection | PubMed |
description | Galleria mellonella larvae are an alternative in vivo model that has been extensively used to study the virulence and pathogenicity of different bacteria due to its practicality and lack of ethical constraints. However, the larvae possess intrinsic autofluorescence that obstructs the use of fluorescent proteins to study bacterial infections, hence better methodologies are needed. Here, we report the construction of a promoter probe vector with bioluminescence expression as well as the optimization of a total bacterial RNA extraction protocol to enhance the monitoring of in vivo infections. By employing the vector to construct different gene promoter fusions, variable gene expression levels were efficiently measured in G. mellonella larvae at various time points during the course of infection and without much manipulation of the larvae. Additionally, our optimized RNA extraction protocol facilitates the study of transcriptional gene levels during an in vivo infection. The proposed methodologies will greatly benefit bacterial infection studies as they can contribute to a better understanding of the in vivo infection processes and pathogen–mammalian host interactions. |
format | Online Article Text |
id | pubmed-7697238 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-76972382020-11-29 Monitoring Gene Expression during a Galleria mellonella Bacterial Infection Moya-Andérico, Laura Admella, Joana Fernandes, Rodrigo Torrents, Eduard Microorganisms Article Galleria mellonella larvae are an alternative in vivo model that has been extensively used to study the virulence and pathogenicity of different bacteria due to its practicality and lack of ethical constraints. However, the larvae possess intrinsic autofluorescence that obstructs the use of fluorescent proteins to study bacterial infections, hence better methodologies are needed. Here, we report the construction of a promoter probe vector with bioluminescence expression as well as the optimization of a total bacterial RNA extraction protocol to enhance the monitoring of in vivo infections. By employing the vector to construct different gene promoter fusions, variable gene expression levels were efficiently measured in G. mellonella larvae at various time points during the course of infection and without much manipulation of the larvae. Additionally, our optimized RNA extraction protocol facilitates the study of transcriptional gene levels during an in vivo infection. The proposed methodologies will greatly benefit bacterial infection studies as they can contribute to a better understanding of the in vivo infection processes and pathogen–mammalian host interactions. MDPI 2020-11-16 /pmc/articles/PMC7697238/ /pubmed/33207842 http://dx.doi.org/10.3390/microorganisms8111798 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Moya-Andérico, Laura Admella, Joana Fernandes, Rodrigo Torrents, Eduard Monitoring Gene Expression during a Galleria mellonella Bacterial Infection |
title | Monitoring Gene Expression during a Galleria mellonella Bacterial Infection |
title_full | Monitoring Gene Expression during a Galleria mellonella Bacterial Infection |
title_fullStr | Monitoring Gene Expression during a Galleria mellonella Bacterial Infection |
title_full_unstemmed | Monitoring Gene Expression during a Galleria mellonella Bacterial Infection |
title_short | Monitoring Gene Expression during a Galleria mellonella Bacterial Infection |
title_sort | monitoring gene expression during a galleria mellonella bacterial infection |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7697238/ https://www.ncbi.nlm.nih.gov/pubmed/33207842 http://dx.doi.org/10.3390/microorganisms8111798 |
work_keys_str_mv | AT moyaandericolaura monitoringgeneexpressionduringagalleriamellonellabacterialinfection AT admellajoana monitoringgeneexpressionduringagalleriamellonellabacterialinfection AT fernandesrodrigo monitoringgeneexpressionduringagalleriamellonellabacterialinfection AT torrentseduard monitoringgeneexpressionduringagalleriamellonellabacterialinfection |