Structural peculiarities of linear megaplasmid, pLMA1, from Micrococcus luteus interfere with pyrosequencing reads assembly.
Different strains of Micrococcus luteus, isolated from high-altitude Argentinean wetlands, were recently reported to harbour the linear plasmids pLMA1, pLMH5 and pLMV7, all of which with 5'-covalently attached terminal proteins. The link between pLMA1 and the host's erythromycin resistance as well as further presumptive qualities prompted us to perform a detailed characterization. When the 454 technology was applied for direct sequencing of gel-purified pLMA1, assembly of the reads was impossible. However, combined Sanger/454 sequencing of cloned pLMA1 fragments, covering altogether 23 kb of the 110-kb spanning plasmid, allowed numerous sequence repeats of varying in lengths to be identified thus rendering an explanation for the above 454 assembly failure. A large number of putative transposase genes were identified as well. Furthermore, a region with five putative iteron sequences is possibly involved in pLMA1 replication.
Wagenknecht
Martin
M
Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, Corrensstr. 3, 48149, Münster, Germany.
Dib
Julián R
JR
Thürmer
Andrea
A
Daniel
Rolf
R
Farías
María E
ME
Meinhardt
Friedhelm
F
D001426:Bacterial Proteins / Q000235:genetics; D004261:DNA Replication; D004269:DNA, Bacterial / Q000737:chemistry* / Q000235:genetics*; D016982:Micrococcus luteus / Q000235:genetics*; D010957:Plasmids; D017422:Sequence Analysis, DNA / Q000379:methods*; D019895:Transposases / Q000235:genetics
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D019895:Transposases