Creation and characterization of glycosyltransferase mutants of trypanosoma brucei

Abstract

The survival strategies of protozoan parasites frequently involve the participation of glycoconjugates. Trypanosoma brucei expresses complex glycoproteins throughout its life cycle and a review of its repertoire of glycosidic linkages suggests a minimum of 38 glycosyltransferase activities. Here we describe a functional characterization workflow in which we create glycosyltransferase null or conditional null mutants in both the bloodstream and procyclic life-cycle forms of the parasite. Subsequently, we characterize the biochemical phenotype of the mutant strains generated and assign precise functions to the genes involved in glycoconjugate biosynthesis and processing in T. brucei. In this way, a comprehensive picture of Â?T. bruceiï?¿ glycosylation associated genes, their specificities and their relationship to similar genes in other organisms can be obtained. © Springer Science+Business Media New York 2013.