Terminal, and telomere-associated proteins of pAL1, a linear plasmid from Arthrobacter nitroguajacolicus Rü61a
Final Report Abstract
A knockout strategy for linear plasmid-borne genes was established, facilitating the knockout of a gene encoding an single-stranded binding protein (ORF39). Though targeted in a similar approach, genes located at the terminus of pAL1, potentially involved in its replication, could not be knocked out, pointing to an essential function. Attempts to identify and clone an internal origin of replication suggest replication exclusively from the ends of the plasmid, a strategy that differs from the hitherto known strategy of other actinomycetes linear plasmids. Refined annotation of ORF100, located in the replication-involved operon, were obtained and compared with other sequenced linear plasmids from actinomycetes. As striking similarities in the genetic organization became evident, our findings are potentially of general significance. Analysis of genetic knockouts involves transition states in which the wild-type and the hybrid plasmid exist in parallel. Since the copy number of pAL1 was found to be rather low, the knockout of essential genes might not be possible at all. From the work directly funded by the project, two publications (one of which is already under revision and the other is in preparation) will originate. As the expertise we obtained throughout the project work contributed to related investigations of Argentinean colleagues, two further publications are listed, in which our support by the DFG is duly acknowledged.
Publications
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(2007) The linear plasmid pAL1 of Arthrobacter nitroguajacolicus Rü61a – termini, transcriptional analysis and copy number. VAAM annual meeting, Osnabrück
Wagenknecht, M., Parschat, K., Fetzner, S., Klassen, R., and Meinhardt, F.
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(2008) Linear plasmid pAL1 of Arthrobacter nitroguajacolicus Rü61a: Terminal protein, and transcriptional analysis of genes potentially involved in replication. VAAM annual meeting, Frankfurt/Main
Wagenknecht, M., Kolkenbrock, S., Meinhardt, F., and Fetzner, S.
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(2009) Large linear plasmids in Micrococcus strains isolated from Argentinean high-altitude wetlands. VAAM annual meeting, Bochum
Dib, J.R., Wagenknecht, M., Farías, M.E., and Meinhardt, F.
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(2009) Novel linear megaplasmid from Brevibacterium sp. isolated from extreme environment. 45th Annual Meeting Argentine Society for Biochemistry and Molecular Biology
Dib, J.R., Wagenknecht, M., Hill, R.T., Farías, M.E., and Meinhardt F.
