Mathiopoulos K D, Powell J D, McCutchan T F
Laboratory of Malaria Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892.
Mol Biol Evol. 1995 Jan;12(1):103-12. doi: 10.1093/oxfordjournals.molbev.a040179.
We introduce here a simple approach for rapidly determining restriction maps for a number of regions of a genome; this involves "anchoring" a map with a rare restriction site (in this case the seldom-cutting EagI) followed by partial digestion of a frequent-cutting enzyme (e.g., Sau 3A). We applied this technology to five species of the Anopheles gambiae complex. In a single Southern blot we obtained about a 15-kb restriction map each for the mtDNA, rRNA gene, and a scnDNA region for each of five species. Phylogenetic analyses of these regions yield trees at odds with the more traditional chromosome inversion-based trees. The value of the approach for systematic purposes is the ease with which several large, independent regions of the genome can be quickly assayed for molecular variation.
我们在此介绍一种快速确定基因组多个区域限制性图谱的简单方法;该方法包括用一个稀有限制性位点(在这种情况下是切割频率低的EagI)“锚定”图谱,然后用一种切割频率高的酶(如Sau 3A)进行部分消化。我们将这项技术应用于冈比亚按蚊复合体的五个物种。在一次Southern杂交中,我们为五个物种中的每一个物种的线粒体DNA、rRNA基因和一个单拷贝核DNA区域分别获得了约15kb的限制性图谱。对这些区域的系统发育分析得出的树状图与更传统的基于染色体倒位的树状图不一致。该方法对于系统研究的价值在于能够轻松快速地检测基因组中几个大的独立区域的分子变异。