Hanotte O, Burke T, Armour J A, Jeffreys A J
Department of Zoology, University of Leicester, United Kingdom.
Genomics. 1991 Apr;9(4):587-97. doi: 10.1016/0888-7543(91)90351-e.
We report here for the first time the large-scale isolation of hypervariable minisatellite DNA sequences from a non-human species, the Indian peafowl (Pavo cristatus). A size-selected genomic DNA fraction, rich in hypervariable minisatellites, was cloned into Charomid 9-36. This library was screened using two multilocus hypervariable probes, 33.6 and 33.15 and also, in a "probe-walking" approach, with five of the peafowl minisatellites initially isolated. Forty-eight positively hybridizing clones were characterized and found to originate from 30 different loci, 18 of which were polymorphic. Five of these variable minisatellite loci were studied further. They all showed Mendelian inheritance. The heterozygosities of these loci were relatively low (range 22-78%) in comparison with those of previously cloned human loci, as expected in view of inbreeding in our semicaptive study population. No new length allele mutations were observed in families and the mean mutation rate per locus is low (less than 0.004, 95% confidence maximum). These loci were also investigated by cross-species hybridization in related taxa. The ability of the probes to detect hypervariable sequences in other species within the same avian family was found to vary, from those probes that are species-specific to those that are apparently general to the family. We also illustrate the potential usefulness of these probes for paternity analysis in a study of sexual selection, and discuss the general application of specific hypervariable probes in behavioral and evolutionary studies.
我们首次在此报告从非人类物种印度孔雀（Pavo cristatus）中大规模分离高变微卫星DNA序列的情况。一个富含高变微卫星的大小选择基因组DNA片段被克隆到Charomid 9-36中。使用两种多位点高变探针33.6和33.15对该文库进行筛选，并且还采用“探针步移”方法，用最初分离的五个孔雀微卫星进行筛选。对48个阳性杂交克隆进行了表征，发现它们来自30个不同的位点，其中18个是多态性的。对其中五个可变微卫星位点进行了进一步研究。它们均显示孟德尔遗传。与先前克隆的人类位点相比，这些位点的杂合度相对较低（范围为22%-78%），鉴于我们半圈养研究群体中的近亲繁殖情况，这是预期的。在家族中未观察到新的长度等位基因突变，每个位点的平均突变率较低（小于0.004，95%置信区间最大值）。还通过跨物种杂交在相关分类群中对这些位点进行了研究。发现探针检测同一鸟类家族中其他物种高变序列的能力各不相同，从物种特异性探针到显然对该家族通用的探针都有。我们还在一项性选择研究中说明了这些探针在亲子鉴定分析中的潜在用途，并讨论了特定高变探针在行为和进化研究中的一般应用。
