Abstract

Summary Aedes albopictus transmits several arboviral infections. In the absence of vaccines, control of mosquito populations is the only strategy to prevent vector‐borne diseases. As part of the search for novel, biological and environmentally friendly strategies for vector control, the isolation of new bacterial species with mosquitocidal activity represents a promising approach. However, new bacterial isolates may be difficult to grow and genetically manipulate. To overcome these limits, here we set up a system allowing the expression of mosquitocidal bacterial toxins in the well‐known genetic background of Bacillus subtilis. As a proof of this concept, the ability of B. subtilis to express individual or combinations of toxins of Bacillus thuringiensis israelensis (Bti) was studied. Different expression systems in which toxin gene expression was driven by IPTG‐inducible, auto‐inducible or toxin gene‐specific promoters were developed. The larvicidal activity of the resulting B. subtilis strains against second‐instar Ae. albopictus larvae allowed studying the activity of individual toxins or the synergistic interaction among Cry and Cyt toxins. The expression systems here presented lay the foundation for a better improved system to be used in the future to characterize the larvicidal activity of toxin genes from new environmental isolates.

Highlights

  • Mosquito-borne viruses and pathogens are responsible for the deadliest diseases, causing more than 700 000 deaths each year (Gates, 2017)

  • The biological control of mosquitoes using bacterial toxins, such as those produced by Bacillus thuringiensis israelensis, is increasingly attracting attention as a possible alternative control strategy in many field settings and the expansion of the toolbox of available toxins active against mosquitoes is desirable (Contreras et al, 2019)

  • The obtained constructs were inserted by double cross-over in the amyE gene of Bacillus subtilis strain PB1831

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Summary

Introduction

Mosquito-borne viruses and pathogens are responsible for the deadliest diseases, causing more than 700 000 deaths each year (Gates, 2017). In order to identify and characterize new toxins from newly isolated mosquitocidal bacteria, it would be useful to create systems allowing the expression of the genes encoding putative new toxins in a well-known genetic background. This strategy would allow investigating the structural and functional properties of the protein of interest using well-studied organisms that are known to be safe for human health, overcoming the limitations of working with environmental isolates that could be difficult to genetically manipulate and possibly toxic for human beings

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