Ferrer María José, Wehrendt Diana Patricia, Bonilla Mariana, Comini Marcelo Alberto, Tellez-Iñón María Teresa, Potenza Mariana
Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, "Dr. Héctor Torres" (INGEBI-CONICET), Buenos Aires, Argentina.
Group Redox Biology of Trypanosomes, Institut Pasteur de Montevideo, Montevideo, Uruguay.
Methods Mol Biol. 2019;1955:105-118. doi: 10.1007/978-1-4939-9148-8_8.
Trypanosomatids are unicellular organisms that colonize a wide diversity of environments and hosts. For instance, Trypanosoma cruzi is a human pathogen responsible for Chagas diseases, while Leishmania tarentolae infects amphibians and became a biotechnological tool suitable for recombinant protein expression. T. cruzi antigens are needed for the development of improved epitope-based methods for diagnosis and treatment of Chagas disease. Molecular cloning for the production of recombinant proteins offers the possibility to obtain T. cruzi antigens at high yield and purity. L. tarentolae appears as the ideal expression host to obtain recombinant T. cruzi antigens with a structure and posttranslational modifications typical of trypanosomatids. In this chapter, we present a protocol for the analytical to mid-scale production of recombinant T. cruzi antigens, using L. tarentolae as expression host (LEXSY inducible system).
锥虫是一类单细胞生物，它们定殖于各种各样的环境和宿主中。例如，克氏锥虫是导致恰加斯病的人类病原体，而塔氏利什曼原虫感染两栖动物，并成为适合重组蛋白表达的生物技术工具。克氏锥虫抗原是开发改进的基于表位的恰加斯病诊断和治疗方法所必需的。通过分子克隆生产重组蛋白为高产率和高纯度地获得克氏锥虫抗原提供了可能性。塔氏利什曼原虫似乎是获得具有锥虫典型结构和翻译后修饰的重组克氏锥虫抗原的理想表达宿主。在本章中，我们介绍了一种以塔氏利什曼原虫为表达宿主（LEXSY诱导系统），用于分析规模至中规模生产重组克氏锥虫抗原的方案。
