Endogenous Linear Plasmids lp28-4 and lp25 are Required for Infectivity and Restriction Protection in the Lyme Disease Spirochete Borrelia mayonii

Single Molecule Real Time (SMRT) sequencing was utilized to map the genome-wide m6A methylation pattern in B. mayonii. Consensus modified motifs were identified in wildtype B. mayonii as well as clonal isolates lacking plasmids that encode predicted methyltransferases. Two conserved m6A modification motifs were identified, and were fully attributable to the presence of specific methyltransferases. In vitro log-phase cultures of Borrelia mayonii isolate MN14-1539 and clones lacking plasmids lp25 and lp28-4 that carry putative restriction modification systems were grown in triplicate. Genomic DNA was isolated from each strain for methylome analysis using PacBio SMRT sequencing.