Southern Hybridization and Restriction Map Analyses of the LdLIP3 locus in the human pathogen Leishmania donovani

Author:  Mary Stromberg and Alison Shakarian
Institution:  Salve Regina University

In the present study Southern hybridization analysis of Leishmania. donovani gDNA was performed to generate a restriction map of the LdLIP3 gene loci and to determine its copy number in the genome. The gDNA was digested with restriction endonucleases and subjected to agarose gel electrophoresis. Restriction fragments were visualized with ethidium bromide staining and subjected to Southern hybridization with a digoxigenin labeled probe of the full length LdLIP3 gene. Results indicated that the gene is present as single copy. Moreover, our data supports the hypothesis that L. donovani possess a gene that encodes a secreted lipase. To date, there are no published studies concerning the characterization of any lipase genes in Leishmania thus this research provides some of the tools necessary to better understand the role of lipases to the biology and malignance of this important human pathogen.