Monitoring Gene Expression during a Galleria mellonella Bacterial Infection

Microorganisms
Laura Moya-AndéricoEduard Torrents

Abstract

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.

References

Dec 15, 2005·Proceedings of the American Thoracic Society·Ruxana T Sadikot, Timothy S Blackwell
Aug 8, 2007·Journal of Invertebrate Pathology·Mariola AndrejkoTeresa Jakubowicz
Sep 4, 2007·Applied and Environmental Microbiology·Christian U RiedelColin Hill
Apr 23, 2009·Microbiology·Olivia L ChampionRichard W Titball
Apr 20, 2011·Infection and Immunity·Britt-Marie Sjöberg, Eduard Torrents
Apr 27, 2012·Applied Microbiology and Biotechnology·Tomás García-CayuelaTeresa Requena
Jul 31, 2012·Applied and Environmental Microbiology·Sabina Leanti La RosaDag Anders Brede
Aug 29, 2013·FEMS Microbiology Letters·Aayushi Jain, Preeti Srivastava
May 9, 2014·Frontiers in Cellular and Infection Microbiology·Eduard Torrents
Feb 6, 2016·Insect Science·Iwona Wojda
Jul 1, 2016·Virulence·Olivia L ChampionRichard W Titball
Aug 3, 2018·Journal of Microbiological Methods·Nathalie ParthuisotJean-Baptiste Ferdy

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Datasets Mentioned

BETA
MW117147

Methods Mentioned

BETA
imaging technique
PCR

Software Mentioned

pETSlux
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ImageJ FIJI
GraphicConverter
GraphPad Prism
NIS

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