Izquierdo Luis, Güther M Lucia S, Ferguson Michael A J
Barcelona Centre for International Health Research, (CRESIB, Hospital Clinic-Universitat de Barcelona), Barcelona, Spain.
Methods Mol Biol. 2013;1022:249-75. doi: 10.1007/978-1-62703-465-4_19.
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 -T. brucei glycosylation associated genes, their specificities and their relationship to similar genes in other organisms can be obtained.
原生动物寄生虫的生存策略通常涉及糖缀合物的参与。布氏锥虫在其整个生命周期中都表达复杂的糖蛋白，对其糖苷键组成的综述表明至少有38种糖基转移酶活性。在这里，我们描述了一种功能表征工作流程，即在寄生虫的血流和前循环生命周期形式中创建糖基转移酶缺失或条件性缺失突变体。随后，我们对所产生的突变菌株的生化表型进行表征，并为参与布氏锥虫糖缀合物生物合成和加工的基因赋予精确功能。通过这种方式，可以获得布氏锥虫糖基化相关基因的全面图谱、它们的特异性以及它们与其他生物体中相似基因的关系。
