AUTHOR=Wadley Taylor , Moon Sun Hee , DeMott Michael S. , Wanchai Visanu , Huang En , Dedon Peter C. , Boysen Gunnar , Nookaew Intawat TITLE=Nanopore Sequencing for Detection and Characterization of Phosphorothioate Modifications in Native DNA Sequences JOURNAL=Frontiers in Microbiology VOLUME=13 YEAR=2022 URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2022.871937 DOI=10.3389/fmicb.2022.871937 ISSN=1664-302X ABSTRACT=
Bacterial DNA is subject to various modifications involved in gene regulation and defense against bacteriophage attacks. Phosphorothioate (PT) modifications are protective modifications in which the non-bridging oxygen in the DNA phosphate backbone is replaced with a sulfur atom. Here, we expand third-generation sequencing techniques to allow for the sequence-specific mapping of DNA modifications by demonstrating the application of Oxford Nanopore Technologies (ONT) and the ELIGOS software package for site-specific detection and characterization of PT modifications. The ONT/ELIGOS platform accurately detected PT modifications in a plasmid carrying synthetic PT modifications. Subsequently, studies were extended to the genome-wide mapping of PT modifications in the