Title : Comparative genomics of Geobacillus species sensitive to TP-84 bacteriophage: identification of the bacterial defense systems
Abstract:
Defense systems are an essential weapon for prokaryotic microorganisms to resist heterologous DNA. Such weapons help them survive the persistent invasion of bacteriophages. Additionally, microbes use various defense systems to filter, control or degrade mobile genetic elements. However, functional defense systems are also a serious obstacle in the genetic engineering of biotechnologically relevant bacterial species, such as Geobacillus stearothermophilus. Due to its unique properties, G. stearothermophilus is a perfect candidate for the construction of a new procaryotic expression system for the production of thermostable recombinant proteins, as well as a bacterial host for a novel phage display system based on thermophilic TP-84 bacteriophage. Unfortunately, Geobacillus stearothermophilus could not be efficiently transformed using recombinant plasmid DNA vectors. To solve this problem, we selected five Geobacillus and Parageobacillus strains sensitive to TP-84 bacteriophage. We isolated genomic DNA from the selected strains and subjected it to DNA sequencing, using three next generation sequencing (NGS) platforms: Oxford Nanopore, Pacific Bioscience, and Illumina. We performed a detailed bioinformatic analysis of the resulting NGS data, compared the obtained complete genome sequences, identified the defense islands, and established patterns of DNA methylation. The identified defense genes will be used as molecular targets for the design and construction of recombinant Geobacillus strains with increased transformation efficiency. This work was financially supported by the National Center for Research and Development (Poland) grant TECHMATSTRATEG2/410747/11/NCBR/2019.
