Isolation and molecular characterization of oligopeptidase B gene of Trypanosoma evansi from Indian dromedaries.

Abstract

The aim of the study was isolation and molecular characterization of oligopeptidase B (opdB) gene of Trypanosoma evansi of camel. Genomic DNA of T. evansi was used to amplify the opdB gene by polymerase chain reaction then amplicon was cloned in a suitable plasmid vector and finally custom sequenced. The desired amplicon of opdB gene of T. evansi was amplified by PCR using gene specific primers. The amplicon of expected size was purified and then ligated to the pGEM-T Easy vector and ligated mixture was transformed into Escherichia coli JM109 strains for cloning. Screening of recombinants was done by restriction enzyme digestion of plasmid DNA. After confirmation of clone of opdB genes the plasmid DNA was sequenced and coding sequences of opdB gene according to the result obtained was of 2092 bp. Tree topology of opdB gene is based on the Neighbor-Joining method and maximum parsimony method with 100% bootstrap values and identified opdB gene sequence showed a close homology with other Trypanosoma and Leishmania spp. gene sequences.