Molecular isolation, cloning and expressions of L– glutaminase encoded gene from the aquatic Streptomyces collected from Persian Gulf

Abstract

L-Glutaminase is a therapeutic enzyme found in various microbial source have been considered in the cancer therapy. Sampling was carried out from the shores of the Persian Gulf. After identifying and performing specific biochemical tests, marine Streptomyces was isolated and DNA extraction was performed. Through the PCR test, the strains of streptomycin with the L-glutamine enzyme gene were identified. The L-gluta gene was positively transmitted to the host bacterium Escherichia coli via a vector and cloned through the TA technique, and the Real Time PCR technique was used to measure the expression of genes in E. coli origami. The software clustalX and Mega5 were used to draw the phylogenetic tree. Out of 12 Streptomyces isolates, 58.3% of isolates were carried L-gluta gene. After cloning the L-glutaminase gene by colony selection (blue / white), the cloned strains were isolated. The real-time PCR test showed a successful expression of the L-gluta gene on the cloned strains. Phylogenetic results with the neighbor joining (NJ) method show that, Streptomyces species with bootstrap values 99% located in a clade which indicated their close relatedness. The results of this study showed that the Persian Gulf is one of the high potential sources with the production of secondary metabolites and useful antimicrobial products that can be used as a useful source of various biological products such as L-glutaminase.