Copy number determination of different derivatives of the streptomycete mini-plasmid pSLG33 |
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Authors: | K Váchalová J Felsberg M Pet?í?ek J Spí?ek P Tichý |
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Institution: | (1) Institute of Microbiology, Academy of Sciences of the Czech Republic, 142 20 Prague, Czech Republic |
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Abstract: | Copy numbers of the streptomycete plasmid vector pRS410 and five other recombinant plasmid derivatives of the original cryptic
streptomycete plasmid pSLG33 were determined using calibrated laser densitometry. DNA preparations, electrophoretically separated
on agarose gels, were stained with ethidium bromide, photographed and the negatives were subsequently scanned in a laser densitometer.
The pSLG33 replicon is very stable, as no effect of the selective pressure was observed. It is a multicopy plasmid with up
to 220 detected copies per chromosome. The use of deletion and/or insertion mutants allowed us to define two regions of the
pSLG33 molecule involved in the control of plasmid replication. |
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