Peña C P, Lander N, Rodríguez E, Crisante G, Añez N, Ramírez J L, Chiurillo M A
Laboratorio de Genética Molecular Dr. Yunis-Turbay, Decanato de Ciencias de la Salud, Universidad Centroccidental Lisandro Alvarado, Barquisimeto, Venezuela.
Acta Trop. 2009 Sep;111(3):255-62. doi: 10.1016/j.actatropica.2009.05.003. Epub 2009 May 9.
Trypanosoma rangeli, a non-pathogenic hemoflagelate that in Central and South America infects humans, shares with Trypanosoma cruzi reservoirs and triatomine vectors, as well as geographical distribution. Recently, we have described in T. rangeli a truncated gene copy belonging to the group II of the trans-sialidase superfamily (TrGP). This superfamily, collectively known in T. cruzi as gp85/TS, includes members that are involved in host cell invasion and infectivity. To confirm the presence of this superfamily in the genome of T. rangeli and obtain a better knowledge of its characteristics, we designed a PCR and RT-PCR cloning strategy to allow sequence analysis of both genomic and transcribed copies. We identified two full-length copies of TrGP, some pseudogenes, and N- and C-terminal sequences of several genes. We also analyzed the expression and cellular localization of these proteins in epimastigote forms of a Venezuelan T. rangeli isolate using polyclonal antibodies made against a recombinant peptide from the N-terminal region of a TrGP member. We confirmed that TrGP is a multigenic family that shares many features with T. cruzi gp85/TS, including the telomeric location of some of its members, and by immunofluorescence analysis that its location is at the surface of the parasite.
克氏锥虫(Trypanosoma rangeli)是一种非致病性血鞭毛虫,在中美洲和南美洲感染人类,它与克氏锥虫(Trypanosoma cruzi)共享宿主和锥蝽传播媒介,以及地理分布。最近,我们在克氏锥虫中描述了一个属于转唾液酸酶超家族(TrGP)第二组的截短基因拷贝。这个超家族在克氏锥虫中统称为gp85/TS,其成员参与宿主细胞入侵和感染性。为了确认这个超家族在克氏锥虫基因组中的存在并更好地了解其特征,我们设计了一种PCR和RT-PCR克隆策略,以便对基因组和转录拷贝进行序列分析。我们鉴定出了两个TrGP全长拷贝、一些假基因以及几个基因的N端和C端序列。我们还使用针对TrGP成员N端区域的重组肽制备的多克隆抗体,分析了这些蛋白在委内瑞拉克氏锥虫分离株的上鞭毛体形式中的表达和细胞定位。我们证实TrGP是一个多基因家族,与克氏锥虫gp85/TS有许多共同特征,包括其一些成员的端粒定位,并通过免疫荧光分析表明其位于寄生虫表面。