Van der Ploeg L H, Cornelissen A W, Barry J D, Borst P
EMBO J. 1984 Dec 20;3(13):3109-15. doi: 10.1002/j.1460-2075.1984.tb02266.x.
We have compared chromosome-sized DNA molecules (molecular karyotypes) of five genera (nine species) of kinetoplastida after cell lysis and deproteinization of DNA in agarose blocks and size fractionation of the intact DNA molecules by pulsed field gradient (PFG) gel electrophoresis. With the possible exception of Trypanosoma vivax and Crithidia fasciculata, all species have at least 20 chromosomes. There are large differences between species in molecular karyotype and in the chromosomal distribution of the genes for alpha- and beta-tubulin, rRNA and the common mini-exon sequence of kinetoplastid mRNAs. In all cases, the rRNA genes are in DNA that is larger than 500 kb. Whereas T. brucei has approximately 100 mini-chromosomes of 50-150 kb, only few are found in T. equiperdum; T. vivax has no DNA smaller than 2000 kb. As all three species exhibit antigenic variation, small chromosomes with telomeric variant surface glycoprotein genes cannot be vital to the mechanism of antigenic variation. The apparent plasticity of kinetoplastid genome composition makes PFG gel electrophoresis a potentially useful tool for taxonomic studies.
我们在琼脂糖块中对DNA进行细胞裂解和脱蛋白处理,并通过脉冲场梯度(PFG)凝胶电泳对完整的DNA分子进行大小分级后,比较了动基体目五个属(九个物种)的染色体大小的DNA分子(分子核型)。除了间日锥虫和 fasciculata 克氏锥虫外,所有物种至少有20条染色体。不同物种在分子核型以及α-和β-微管蛋白、rRNA基因和动基体目mRNA的常见小外显子序列的染色体分布上存在很大差异。在所有情况下,rRNA基因都存在于大于500 kb的DNA中。布氏锥虫有大约100条50 - 150 kb的小染色体,而在马媾疫锥虫中只发现少数;间日锥虫没有小于2000 kb的DNA。由于这三个物种都表现出抗原变异,带有端粒变异表面糖蛋白基因的小染色体对于抗原变异机制可能并非至关重要。动基体目基因组组成的明显可塑性使得PFG凝胶电泳成为分类学研究中一种潜在有用的工具。