Data from: A self-transmissible plasmid from a hyperthermophile that facilitates genetic modification of diverse Archaea

Conjugative plasmids (CP) are self-transmissible mobile genetic elements
which transfer DNA between host cells via Type IV Secretion Systems
(T4SS). While T4SS-mediated conjugation has been well-studied in bacteria,
information is sparse in Archaea and known representatives exist only in
the Sulfolobales order of Crenarchaeota. Here we present the first
self-transmissible plasmid identified in a Euryarchaeon, Thermococcus sp.
33-3. The 103 kbp plasmid, pT33-3, is seen in CRISPR-spacers throughout
the Thermococcales order. We demonstrate that pT33-3 is a bona fide
conjugative plasmid that requires cell-to-cell contact, and is dependent
upon canonical, plasmid-encoded T4SS-like genes. Under laboratory
conditions pT33-3 transfers to various Thermococcales, and transconjugants
propagate at 100°C. Using pT33-3, we developed a genetic toolkit which
allows modification of phylogenetically diverse Archaeal genomes. We
demonstrate pT33-3–mediated plasmid mobilization and subsequent targeted
genome modification in previously untransformable Thermococcales species,
and extend this process to interphylum transfer to a Crenarchaeon.