Van Den Bussche R A, Honeycutt R L, Baker R J
Department of Biological Sciences, Texas Tech University, Lubbock 79409.
Genetica. 1992;87(3):141-9. doi: 10.1007/BF00240553.
Constitutive heterochromatin of a karyotypically conserved species of harvest mouse was compared to that of three karyotypically derived species of harvest mice by examining banding patterns produced on metaphase patterns produced by two of these restriction endonucleases (EcoRI and MboI) were compared to published G- and C-banded karyotypes and in situ hybridization of a satellite DNA repeat for these taxa. The third restriction endonuclease (PstI) did not produce a detectable pattern of digestion. For the most part, patterns produced by EcoRI and MboI can be related to C-banded chromosomes and in situ hybridization of satellite DNA sequences. Moreover, digestion with EcoRI reveals bands not apparent with these other techniques, suggesting that restriction endonuclease digestion of metaphase chromosomes may provide additional insight into the structure and organization of metaphase chromosomes. The patterns produced by restriction endonuclease digestion are compatible with the chromosomal evolution of these taxa, documenting that in the highly derived taxa not only are the chromosomes rearranged but the abundance of certain sequences is highly variable. However, technical variation and difficulty in producing consistent results even on a single slide with some restriction endonucleases documents the problems associated with this method.
通过检查由两种限制性内切酶(EcoRI和MboI)在中期模式上产生的带型,将一种核型保守的收获鼠物种的组成型异染色质与三种核型衍生的收获鼠物种的组成型异染色质进行了比较。将这些限制性内切酶产生的带型与已发表的G带和C带核型以及这些分类群的卫星DNA重复序列的原位杂交结果进行了比较。第三种限制性内切酶(PstI)未产生可检测到的消化模式。在大多数情况下,EcoRI和MboI产生的带型可与C带染色体以及卫星DNA序列原位杂交结果相关联。此外,用EcoRI消化可揭示出其他技术无法显现的带型,这表明对中期染色体进行限制性内切酶消化可能会为中期染色体的结构和组织提供更多见解。限制性内切酶消化产生的带型与这些分类群的染色体进化情况相符,证明在高度衍生的分类群中,不仅染色体发生了重排,而且某些序列的丰度也高度可变。然而,技术差异以及即使在同一张载玻片上使用某些限制性内切酶也难以产生一致结果,这证明了该方法存在的问题。