McCulloch Richard, Vassella Erik, Burton Peter, Boshart Michael, Barry J David
Welcome Centre for Molecular Parasitology, University of Glasgow, United Kingdom.
Methods Mol Biol. 2004;262:53-86. doi: 10.1385/1-59259-761-0:053.
African trypanosomes, such as Trypanosoma brucei, are protozoan parasites of mammals that were first described over 100 hundred years ago. They have long been the subjects of biological investigation, which has yielded insights into a number of fundamental, as well as novel, cellular processes in all organisms. In the last decade or so, genetic manipulation of trypanosomes has become possible through DNA transformation, allowing yet more detailed analysis of the biology of the parasite. One facet of this is that DNA transformation has itself been used as an assay for recombination and will undoubtedly lead to further genetic approaches to examine this process. Here we describe protocols for DNA transformation of Trypanosoma brucei, including two different life cycle stages and two different strain types that are distinguished by morphological and developmental criteria. We consider the application of transformation to recombination, as well as the uses of transforming the different life cycle stages and strain types.
非洲锥虫,如布氏锥虫,是哺乳动物的原生动物寄生虫,早在100多年前就被首次描述。长期以来,它们一直是生物学研究的对象,这使人们对所有生物体中的许多基本以及新颖的细胞过程有了深入了解。在过去十年左右的时间里,通过DNA转化实现了对锥虫的基因操作,从而能够对该寄生虫的生物学进行更详细的分析。其中一个方面是,DNA转化本身已被用作重组分析,无疑将导致进一步研究这一过程的基因方法。在这里,我们描述了布氏锥虫DNA转化的方案,包括两个不同的生命周期阶段和两种通过形态和发育标准区分的不同菌株类型。我们考虑了转化在重组中的应用,以及不同生命周期阶段和菌株类型转化的用途。