Molecular cloning and characterization of a Leishmania donovani alpha-tubulin gene

We have isolated a cDNA for an alpha-tubulin mRNA from L. donovani promastigotes and determined its complete nucleotide sequence. Both nucleotide and deduced amino acid sequence analysis of this cDNA showed significant similarity with a previously reported, partial sequence of an L. enriettii alpha-tubulin and the complete sequence of human alpha-tubulin. Further, the in vitro translated L. donovani alpha-tubulin gene product was specifically immunoprecipitated with a monoclonal antibody against human alpha-tubulin. Northern blot analysis revealed that there was little change in the expression of the L. donovani alpha-tubulin RNA during parasite differentiation from promastigote to the in vitro grown "amastigote" form. Southern blot analysis revealed a simple genomic organization for the L. donovani alpha-tubulin gene with more than one copy of the alpha-tubulin gene in the parasite genome. To our knowledge, this is the first complete sequence of an alpha-tubulin for Leishmania to be reported in the literature.