Molecular characterization of trans-sialidase gene of Trypanosoma evansi isolated from Indian dromedaries

Abstract

The present study was carried out to isolate the Trans-sialidase (ts) gene of Trypanosoma evansi by polymerase chain reaction, clone the amplicons in a suitable bacterial plasmid vector and characterise the gene through sequencing. The desired amplicon of ts gene of T. evansi was amplified by PCR using gene specific primers and identified on the basis of size of the gene. The amplicon of expected size was purified from the 1% low melting agarose gel. The DNA fragment of interest was then ligated to the pGEM-T Easy vector and ligated mixture was transformed into Escherichia coli JM109 strains for cloning. After cloning, screening of recombinants was done by Restriction Enzyme digestion of plasmid DNA and by colony PCR. After confirmation of clone, the plasmid DNA was sequenced and coding sequence of ts gene according to the results obtained was of 2241 bp. Tree topology of ts gene is based on the Neighbour-Joining method with 100% bootstrap values and identified ts gene sequence showed a close homology with other Trypanosoma spp. gene sequences.