Kian Danielle, Lancheros César Armando Contreras, Damiani Igor Alexandre Campos, Fernandes Tamiris Zanforlin Olmos, Pinge-Filho Phileno, Santos Márcia Regina Machado dos, Silveira José Franco da, Nakamura Celso Vataru, Silva João Santana da, Yamada-Ogatta Sueli Fumie, Yamauchi Lucy Megumi
Departamento de Microbiologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina, PR, Brazil.
Departamento de Microbiologia, Imunobiologia e Parasitologia, Universidade Federal de São Paulo, São Paulo, SP, Brazil.
Korean J Parasitol. 2015 Aug;53(4):483-8. doi: 10.3347/kjp.2015.53.4.483. Epub 2015 Aug 25.
This report describes the molecular characterization of the Tc8.2 gene of Trypanosoma cruzi. Both the Tc8.2 gene and its encoded protein were analyzed by bioinformatics, while Northern blot and RT-PCR were used for the transcripts. Besides, immunolocalization of recombinant protein was done by immunofluorescence and electron microscopy. Analysis indicated the presence of a single copy of Tc8.2 in the T. cruzi genome and 2-different sized transcripts in epimastigotes/amastigotes and trypomastigotes. Immunoblotting showed 70 and 80 kDa polypeptides in epimastigotes and trypomastigotes, respectively, and a differential pattern of immunolocalization. Overall, the results suggest that Tc8.2 is differentially expressed during the T. cruzi life cycle.
本报告描述了克氏锥虫Tc8.2基因的分子特征。通过生物信息学分析了Tc8.2基因及其编码蛋白,同时利用Northern印迹和RT-PCR分析转录本。此外,通过免疫荧光和电子显微镜对重组蛋白进行免疫定位。分析表明,克氏锥虫基因组中存在单个拷贝的Tc8.2,在无鞭毛体/前鞭毛体和锥鞭毛体中有两种不同大小的转录本。免疫印迹显示,前鞭毛体和锥鞭毛体中分别存在70 kDa和80 kDa的多肽,且免疫定位模式不同。总体而言,结果表明Tc8.2在克氏锥虫生命周期中差异表达。