European Journal of Experimental Biology Open Access

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Molecular characterization of Pseudomonas sp. isolated from milk samples by using RAPD-PCR

Ahmed Majid Hameed, Sudhakar Malla and R. Senthil Kumar

Degradation of milk and its products is one of the major breakthroughs what many dairy based industries are facing. Despite of recent advances in antibiotic therapy, the industries are still facing the issue seriously. The spoilage of the milk and reduction in the shelf life of processed milk takes place through various enzymatic activities associated with the contamination of dairy products by Pseudomonas spp. High diversity might be one of the possible reason for a strain to adapt to the new environment, which is useful for its propagation and domestication of wild species. In this study, we attempted to determine the use of RAPD-PCR as an effective tool to detect genetic variation between P. aeuroginosa species isolated from the different milk samples. Milk samples were obtained from different areas both from the local field and dairy farms. The samples were screened for the pseudomonas and DNA was extracted. The DNA thus obtained was used for RAPD PCR using the OPZ-8 and OPD-20 primers to reveal the genetic diversity among the variants. The cluster analysis based on the RAPD analysis was done separately for both OPD-20 and OPZ-8 and in case of OPZ-8 cluster analysis, Major group A was divided into two sub groups, first subgroup containing only four accessions, namely, 13, 10, 7 & 8 and Second subgroup containing nine accessions as 6,4,514,3,9,2,11 & 12. In case of the primer OPZ-8 cluster analysis, Major group A was divided into only one sub group containing only four accessions, namely 7, 5,8 & 1. Group B containing ten accessions, namely 6, 3, 4, 14, 11, 9, 12, 2 & 10. All bands showed polymorphism and there was no monomorphic band observed common to all the samples. The number of bands and banding pattern were variable depending upon the primer and type of species tested.