Isolation, amplification and sequence analysis of the gene encoding Enolase from lymphatic filarial parasite Brugia malayi

Lymphatic filariasis, caused by Brugia malayi, commonly known as elephantiasis, is a neglected tropical disease. No vaccines are available for the prevention of filarial infections. A number of pathogenic organisms including filarial parasites display specialized proteins on their cell surface to assist in invasion. One of the best characterized is the glycolytic enzyme enolase. Enolase represents a multifunctional protein involved in basic energy metabolism in pathogens. In the present study, gene encoding enolase of B. malayi was isolated, amplified and identified by sequencing. The amplification and sequencing was done using specific primers. The primers were designed based on the complete genome contig sequence of B. malayi to amplify the cDNA of enolase. The full length cDNA of this gene from B. malayi was obtained by overlapping the sequences of both amplification products using BioEdit version. The results showed that the full length cDNA comprised of 1314 bp. The gene encoding enolase from B. malayi (BmEno) was identified by BLAST result. The sequence of the B. malayi enolase was found to be identical to that of the B. malayi partial coding sequences. The complete coding sequence of B. malayi enolase was submitted to GenBank and accession number (KF830990.1) was obtained. Phylogenetic analysis of B. malayi enolase revealed the occurrence of homology with closely related filarial parasites. Further studies are being carried out to clone and express the enolase gene in the expression vector to study its enzyme activity for therapeutic potential.