Praveen Pal Balgir, Puja Bhatia, Pushpinder Kaur and Vipin Kumar* Department of Biotechnology, Punjabi University, Patiala-147002, Punjab, India.
*Directorate of Research, SVPUA&T, Meerut, 250110 (UP), INDIA, E-mail of corresponding author: puja17bt@gmail.com
Abstract: The DNA-intercalating agents, ethidium bromide and ascorbic acid (vitamin C), were used to eliminate plasmid DNA from Pediococcus acidilactici MTCC 5101, a lactic acid bacteria. The strain was grown in the presence of 0.03 mM ethidium bromide or 1 mM ascorbate for 18 hours at 42°C, which resulted in the loss of its ability to produce pediocin, a plasmid-specified trait. Agarose gel electrophoresis of plasmid DNA and spot-on-lawn assay showed concomitant loss of an 8.9 kb plasmid pCP289 coding for pediocin from these colonies. Since ascorbic acid is a readily available, shows less effect on cell viability, nonhazardous and more efficient compound in contrast to ethidium bromide, the possibility of its use in determining plasmidencoded traits in food grade lactic acid bacteria is also proposed.
Keywords: Curing, ethidium bromide, ascorbic acid, Pediococcus acidilactici MTCC 5101
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