Genetic engineering of marine cyanophages reveals integration but not lysogeny in T7-like cyanophages
Name
s41396-021-01085-8.pdf
Description
Published version
Size
2.74 MB
Format
Adobe PDF
Checksum (MD5)
1e2e1043001931511fb5cb518e92c0c3
Author(s)
Shitrit, Dror
Hackl, Thomas
Laurenceau, Raphael
Raho, Nicolas
Carlson, Michael CG
Sabehi, Gazalah
Schwartz, Daniel A
Chisholm, Sallie W
Lindell, Debbie
Date Issued
August 2021
Journal
ISME Journal
Publisher
Springer Science and Business Media LLC
Citation
Shitrit, D., Hackl, T., Laurenceau, R. et al. Genetic engineering of marine cyanophages reveals integration but not lysogeny in T7-like cyanophages. ISME J (2021)
Version
Final published version
Abstract
Marine cyanobacteria of the genera Synechococcus and Prochlorococcus are the most abundant photosynthetic organisms on earth, spanning vast regions of the oceans and contributing significantly to global primary production. Their viruses (cyanophages) greatly influence cyanobacterial ecology and evolution. Although many cyanophage genomes have been sequenced, insight into the functional role of cyanophage genes is limited by the lack of a cyanophage genetic engineering system. Here, we describe a simple, generalizable method for genetic engineering of cyanophages from multiple families, that we named REEP for REcombination, Enrichment and PCR screening. This method enables direct investigation of key cyanophage genes, and its simplicity makes it adaptable to other ecologically relevant host-virus systems. T7-like cyanophages often carry integrase genes and attachment sites, yet exhibit lytic infection dynamics. Here, using REEP, we investigated their ability to integrate and maintain a lysogenic life cycle. We found that these cyanophages integrate into the host genome and that the integrase and attachment site are required for integration. However, stable lysogens did not form. The frequency of integration was found to be low in both lab cultures and the oceans. These findings suggest that T7-like cyanophage integration is transient and is not part of a classical lysogenic cycle.
MIT Department
Massachusetts Institute of Technology. Department of Civil and Environmental Engineering
Massachusetts Institute of Technology. Department of Biology
Terms of Use
Creative Commons Attribution 4.0 International license
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1038/s41396-021-01085-8
