Structural peculiarities of linear megaplasmid, pLMA1, from Micrococcus luteus interfere with pyrosequencing reads assembly.

Abstract:

: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.

journal_name

Biotechnol Lett

journal_title

Biotechnology letters

authors

Wagenknecht M,Dib JR,Thürmer A,Daniel R,Farías ME,Meinhardt F

doi

10.1007/s10529-010-0357-y

subject

Has Abstract

pub_date

2010-12-01 00:00:00

pages

1853-62

issue

12

eissn

0141-5492

issn

1573-6776

journal_volume

32

pub_type

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