Conditions to enhance transfection in Streptococcus pneumoniae

Abstract

Enhancement of transfection in the wild-type strain of Streptococcus pneumoniae, which contains normal levels of nucleases, can be achieved by a careful control of the components of the medium. Protection of the donor DNA against the degradation by the nucleases present at the surface of the cell during the adsorption to the physiologically competent bacteria is a necessary condition to obtain an enhanced transfection. Using [3H]thymidine-labeled Dp-4 DNA we have found that maximal levels of DNA uptake and a remarkable stimulation in genetic transfection were obtained in the presence of calcium ions. Concentrations of magnesium ions higher than 2 mM stimulated the extracellular degradation of the donor DNA and inhibited transfection.