Abstract
The covalently closed (CC) monomer form of plasmid pMV158 was found to transform pneumococcus (Streptococcus pneumoniae) and to do so with two-hit kinetics. The evidence came from analysis of the behavior of the transforming activity in fractions from preparative gel electrophoresis. Activity in the first major peak to elute (i) co-eluted with monomer CC as detected on analytical gels, (ii) banded as CC in dye-buoyancy gradients, (iii) sedimented with the velocity expected for monomer CC, and (iv) gave two-hit kinetics as functions of both concentration and time of exposure of the cells to DNA. A second major peak of activity behaved physically as though mostly due to dimer CC forms and gave single-hit response curves. Because almost no dimer was detectable optically on analytical gels of starting preparations, its specific activity was high relative to that of the monomers.
MeSH Terms
Centrifugation, Density Gradient
DNA, Bacterial/genetics,isolation & purification
DNA, Circular/genetics,isolation & purification
Electrophoresis, Agar Gel
Kinetics
Plasmids
Streptococcus pneumoniae/genetics
Transformation, Genetic
Chemicals
DNA, Bacterial
DNA, Circular
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Saunders C W
Guild W R
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