Toaldo C B, Steindel M, Sousa M A, Tavares C C
Departamento de Biologia Celular e Genética, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ, 20550-013, Brasil.
Mem Inst Oswaldo Cruz. 2001 Jan;96(1):113-21. doi: 10.1590/s0074-02762001000100013.
The molecular karyotype of nine Trypanosoma rangeli strains was analyzed by contour-clamped homogeneous electric field electrophoresis, followed by the chromosomal localization of beta-tubulin, cysteine proteinase, 70 kDa heat shock protein (hsp 70) and actin genes. The T. rangeli strains were isolated from either insects or mammals from El Salvador, Honduras, Venezuela, Colombia, Panama and southern Brazil. Also, T. cruzi CL-Brener clone was included for comparison. Despite the great similarity observed among strains from Brazil, the molecular karyotype of all T. rangeli strains analyzed revealed extensive chromosome polymorphism. In addition, it was possible to distinguish T. rangeli from T. cruzi by the chromosomal DNA electrophoresis pattern. The localization of beta-tubulin genes revealed differences among T. rangeli strains and confirmed the similarity between the isolates from Brazil. Hybridization assays using probes directed to the cysteine proteinase, hsp 70 and actin genes discriminated T. rangeli from T. cruzi, proving that these genes are useful molecular markers for the differential diagnosis between these two species. Numerical analysis based on the molecular karyotype data revealed a high degree of polymorphism among T. rangeli strains isolated from southern Brazil and strains isolated from Central and the northern South America. The T. cruzi reference strain was not clustered with any T. rangeli strain.
采用轮廓夹钳均匀电场电泳分析了9株克氏锥虫的分子核型，随后对β-微管蛋白、半胱氨酸蛋白酶、70 kDa热休克蛋白（hsp 70）和肌动蛋白基因进行了染色体定位。这些克氏锥虫菌株分别从萨尔瓦多、洪都拉斯、委内瑞拉、哥伦比亚、巴拿马和巴西南部的昆虫或哺乳动物中分离得到。此外，还纳入了克氏锥虫CL-Brener克隆株用于比较。尽管在来自巴西的菌株之间观察到高度相似性，但所有分析的克氏锥虫菌株的分子核型都显示出广泛的染色体多态性。此外，通过染色体DNA电泳图谱可以区分克氏锥虫和枯氏锥虫。β-微管蛋白基因的定位揭示了克氏锥虫菌株之间的差异，并证实了来自巴西的分离株之间的相似性。使用针对半胱氨酸蛋白酶、hsp 70和肌动蛋白基因的探针进行杂交分析，区分了克氏锥虫和枯氏锥虫，证明这些基因是这两个物种鉴别诊断的有用分子标记。基于分子核型数据的数值分析显示，从巴西南部分离的克氏锥虫菌株与从中南美洲中部和北部分离的菌株之间存在高度多态性。克氏锥虫参考菌株未与任何克氏锥虫菌株聚类。
