Barker D C, Butcher J
Trans R Soc Trop Med Hyg. 1983;77(3):285-97. doi: 10.1016/0035-9203(83)90146-3.
Kinetoplast DNA (kDNA), initially characterized by buoyant density, from ten reference isolates of the Leishmania braziliensis and L. mexicana complexes has been radio-actively labelled and used as hybridization probes. Filters containing endonuclease digested, electrophoresed, Southern transferred fragments of kDNA from reference and other isolates sent to us for DNA typing have been tested for kDNA sequence homology. We record a complete lack of sequence homology between kDNA of any isolate of the L. braziliensis complex and kDNA of any isolate of the L. mexicana complex. L. b. braziliensis, L. b. guyanensis and L. b. panamensis have kDNA sequences in common with each other and with a number of test isolates from Brazil, Panama, Venezuela and Peru. L. b. panamensis (1.695 g/ml) can be separated from L. b. braziliensis or L. b. guyanensis (1.691-1.693 g/ml) by differences in buoyant density of kDNA. L. m. mexicana and L. m. pifanoi have kDNA sequences in common with each other but kDNA of L. m. amazonensis has insignificant homology with kDNA of other reference isolates of the L. mexicana complex. We conclude that the kDNAs of species of the L. mexicana complex are sufficiently different from kDNA of species of the L. braziliensis complex to make kDNA sequence homology identification a feasible proposition.
最初通过浮力密度进行表征的动基体DNA(kDNA),取自巴西利什曼原虫和墨西哥利什曼原虫复合体的十个参考分离株,已进行放射性标记并用作杂交探针。我们对含有经核酸内切酶消化、电泳、Southern转移的参考分离株和其他送来进行DNA分型的分离株的kDNA片段的滤膜,检测了kDNA序列同源性。我们发现,巴西利什曼原虫复合体的任何分离株的kDNA与墨西哥利什曼原虫复合体的任何分离株的kDNA之间完全缺乏序列同源性。巴西利什曼原虫巴西亚种、圭亚那利什曼原虫和巴拿马利什曼原虫彼此之间以及与来自巴西、巴拿马、委内瑞拉和秘鲁的一些测试分离株具有共同的kDNA序列。巴拿马利什曼原虫(1.695 g/ml)可通过kDNA浮力密度的差异与巴西利什曼原虫巴西亚种或圭亚那利什曼原虫(1.691 - 1.693 g/ml)区分开来。墨西哥利什曼原虫墨西哥亚种和皮氏利什曼原虫具有共同的kDNA序列,但亚马逊利什曼原虫的kDNA与墨西哥利什曼原虫复合体的其他参考分离株的kDNA具有不显著的同源性。我们得出结论,墨西哥利什曼原虫复合体各物种的kDNA与巴西利什曼原虫复合体各物种的kDNA差异足够大,使得kDNA序列同源性鉴定成为一个可行的方案。