Plasmid DNA was isolated from lactic acid bacteria (LAB) in order to check for plasmid mediated antibiotic resistance. Attempts were made to transfer plasmid with antibiotic resistance to plasmid deficient probiotic strain – Lactobacillus helveticus MTCC 5463. Total 31 LAB isolates were tested for the presence of plasmid DNA using Ultra clean mini plasmid DNA isolation kit (Molbio Laboratories Inc.). After plasmid isolation the samples were run on agarose gel (0.8%) and the gel was observed under UV Transilluminator for the bands of plasmid DNA. The samples showed the presences of plasmid were checked for the plasmid DNA concentration and purity using nanodrop instrument. LAB isolates which showed good plasmid DNA concentration and purity was further taken for identifying plasmid mediated antibiotic resistance. The isolated plasmid was then transferred into plasmid deficient probiotic Lb. helveticus MTCC 5463 using electroporator. Successful transfer of plasmid to Lb. helveticus MTCC 5463 was checked by isolation of plasmid from transformed cells and then developed antibiotic resistance was determined by disc diffusion assay. Out of 31, only 7 LAB isolates were containing plasmid. The highest concentration and purity of plasmid DNA was found in Lb rhamnosus 52 (155.4/2 μl sample). Then Lb rhamnosus 52 plasmid was electroporated successfully in probiotic Lb. helveticus MTCC 5463. It was found that antibiotic resistance against many antibiotics was significantly increased in electroporated cells of Lb. helveticus MTCC 5463 compared to control cells.