Molecular detection of anticancer protein Azurin in native Pseudomonas isolates

Abstract

Azurin is a potent anticancer protein which has the ability to interfere in tumor growth. The purpose of this study was to isolate azurin producing native Pseudomonas isolates along with the confirmation of azurin presence using molecular techniques. In this study, azurin presence was Azurin is a potent anticancer protein which has the ability to interfere in tumor growth. The purpose of this study was to isolate azurin producing native Pseudomonas spp along with the confirmation of azurin presence using molecular techniques. Primarily, the isolates were characterized morphologically and identified biochemically as Pseudomonas spp. Presence of azurin gene was tested for 25 environmental (soil) isolates using specific oligonucleotide primers (AZU-F and AZU-R) in a gradient PCR. The extracted total cellular protein was precipitated by 60% ammonium sulfate followed by dialysis and partially purified azurin using SDS-PAGE. Out of 25 isolates, 10 showed the presence of azurin gene by amplifying a single 545 bp DNA fragment in PCR. Total cellular partially purified protein extracted from 10 azurin producing isolates selected from PCR fractionated by SDS PAGE produced patterns with a wide range of discrete bands with the inclusion of 14 kDa Molecular weight. Presence of azurin gene was confirmed by PCR, and partially purified azurin protein was detected by SDS-PAGE according to specific molecular weight. Detection of high yield azurin producing microbial strains can lead us to the development of anticancer drugs.