Characterization of the effectors required for stable inheritance of Streptococcus pyogenes pSM19035-derived plasmids in Bacillus subtilis |
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Authors: | Piotr Ceglowski Alexander Boitsov Natalia Karamyan Sunghee Chai Juan C. Alonso |
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Affiliation: | (1) Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, PL-02-106 Warsaw, Poland;(2) Max-Planck-Institut für molekulare Genetik, Ihnestrasse 73, D-14195 Berlin, Germany;(3) Present address: Department of Biophysics, Saint Petersburg State Technical University, Saint Petersburg, Russia |
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Abstract: | The low-copy-number and broad-host-range pSM19035-derived plasmid pBT233 is stably inherited in Bacillus subtilis cells. Two distinct regions, segA and segB, enhance the segregational stability of the plasmid. Both regions function in a replicon-independent manner. The maximization of random plasmid segregation is accomplished by the recombination proficiency of the host or the presence of the pBT233 segA region. The segA region contains two open reading frames (or) [ and ]. Inactivation or deletion of or results in SegA– plasmids. Better than random segregation requires an active segB region. The segB region contains two ors (or and or). Inactivation of either of the orfs does not lead to an increase in cell death, but or– plasmids are randomly segregated. These results suggest that pBT233 stabilization relies on a complex system involving resolution of plasmid oligomers (segA) and on the function(s) encoded by the segB region. |
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Keywords: | Partition Site-specific recombination Homologous recombination Rolling circle and theta replication |
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