Molecular Detection of Azurin: A Powerful Anticancer Protein from Native Pseudomonas Isolates

Abstract

Aim: The present study was carried out to isolate azurin producing native Pseudomonas aeruginosa strains and confirm its presence using molecular techniques. Study Design: Presence of azurin detection by molecular techniques such as PCR and SDS PAGE. Place and Duration of the Study: Department of Life Sciences, University of Calicut, Kerala, India, between September 2013 to December 2014. Methodology: Conventional microbiological techniques were used to isolate several Pseudomonas aeruginosa from environmental sources. The presence of azurin gene was confirmed by PCR. The extracted total cellular protein was precipitated by 70% ammonium sulfate followed by dialysis and partially purified azurin fractionated using SDS-PAGE. Results: Out of 10 samples selected for this study 8 shows the presence of azurin gene using specific oligonucleotide primers in a gradient PCR, amplifying a single 545 bp DNA fragment characteristic of azurin gene. The extracted total cellular protein analysed using SDS-Polyacrylamide gel electrophoresis, wide range of bands were present including a band corresponding to 14 KD Molecular weight in all 8 samples selected from PCR. Conclusion: Presence of azurin gene was confirmed by PCR and partially purified protein fractionated in 15% SDS PAGE. Further study of purification and cytotoxic effect of azurin is required.