4.4 Article

Genetic Modification of Sodalis Species by DNA Transduction

Journal

MSPHERE
Volume 6, Issue 1, Pages -

Publisher

AMER SOC MICROBIOLOGY
DOI: 10.1128/mSphere.01331-20

Keywords

Sodalis praecaptivus; Sodalis glossinidius; insect endosymbiont; symbiont; transformation; transduction; genetic modification; plasmid transfer; transposition; bacteriophage P1; gene disruption; mutation; paratransgenesis; bacteriophage transduction; symbiosis

Categories

Funding

  1. National Institutes of Health [AI148774]
  2. Pennsylvania State University College of Medicine

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Bacteriophages play central roles in microbial ecology and evolution by promoting horizontal gene transfer among bacterial species. Bacteriophage P1 has been widely exploited as an experimental tool for the genetic study of bacteria, with the ability to infect, lysogenize, and promote transduction in members of the bacterial genus Sodalis. This research suggests that phage P1 could be used to genetically modify insect endosymbionts without the need for culturing these organisms.
Bacteriophages (phages) are ubiquitous in nature. These viruses play a number of central roles in microbial ecology and evolution by, for instance, promoting horizontal gene transfer (HGT) among bacterial species. The ability of phages to mediate HGT through transduction has been widely exploited as an experimental tool for the genetic study of bacteria. As such, bacteriophage P1 represents a prototypical generalized transducing phage with a broad host range that has been extensively employed in the genetic manipulation of Escherichia coli and a number of other model bacterial species. Here we demonstrate that P1 is capable of infecting, lysogenizing, and promoting transduction in members of the bacterial genus Sodalis, including the maternally inherited insect endosymbiont Sodalis glossinidius. While establishing new tools for the genetic study of these bacterial species, our results suggest that P1 may be used to deliver DNA to many Gram-negative endosymbionts in their insect host, thereby circumventing a culturing requirement to genetically manipulate these organisms. MPORTANCE A large number of economically important insects maintain intimate associations with maternally inherited endosymbiotic bacteria. Due to the inherent nature of these associations, insect endosymbionts cannot be usually isolated in pure culture or genetically manipulated. Here we use a broad-host-range bacteriophage to deliver exogenous DNA to an insect endosymbiont and a closely related free-living species. Our results suggest that broad-host-range bacteriophages can be used to genetically alter insect endosymbionts in their insect host and, as a result, bypass a culturing requirement to genetically alter these bacteria.

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