Blake R D, Wang J Z, Beauregard L
Department of Biochemistry, Microbiology and Molecular Biology, University of Maine, Orono, ME 04469-5735, USA.
J Mol Evol. 1997 May;44(5):509-20. doi: 10.1007/pl00006175.
High-resolution derivative melting was used to obtain detailed distributions of local (G + C) contents in a number of ruminant DNAs. Profiles over low (G + C) regions [20-36% (G + C)] are congruent for all ruminants. This region represents 45-50% of the nuclear DNA content and primarily contains intergenic and intron sequences. The high (G + C) region, where most coding sequences are found [38-68% (G + C)], is marked by satellite bands denoting the presence of transcriptionally inert, tandemly repetitive sequence families. These bands can be analyzed for the abundance, base composition, and sequence divergence of satellite families with relatively high precision. Band patterns are unique to each species; even closely related species can be readily distinguished by their base distribution profiles. Variations in nuclear DNA contents in ruminants, determined by flow cytometry, are primarily due to variations in abundances of these repetitive sequence families. Thus, A. alces (moose) is found to have 8.85 +/- 0.2 pg DNA/cell, 25% more than the average in ruminants, while the base distribution curve indicates the presence of an unusually abundant satellite of 52.6% (G + C). The size (1 kb) and sequence of this satellite corresponds to satellite-I of other cervids, and in consequence it is designated Alces-I. The sequence of a cloned repeat of Alces-I has a length of 968 bp, a (G + C) content of 52.6%, and contributes 35%, or almost 3 million copies to the nuclear DNA, exceeding by approximately 300% the average array size of this repeat family in related cervids. In situ hybridization indicates the repeat is distributed throughout centromeric regions of all 62 acrocentric autosomes. Alces-I has much greater-than-expected numbers of GG, GA, and AG and far fewer numbers of TA and CG duplets, characteristics of all tandem repeats. The sequence is judged to be orthologous with satellite-I sequences from Rangifer tarandus (caribou), Capreolus capreolus (roe deer), Muntiacus muntjac (Chinese muntjac) and Muntiacus reevesi (Indian muntjac), as well as Antilocapra americana (pronghorn), and the bovids Bos taurus and Ovis aries. A tentative tree for the five cervids is in excellent agreement with one proposed on the basis of morphological characteristics. Differences from a consensus sequence indicate transversions exceed transitions by almost twofold, suggesting that substitutions occur randomly, or nearly so.
高分辨率衍生熔解用于获取多种反刍动物DNA中局部(G + C)含量的详细分布。所有反刍动物在低(G + C)区域[20 - 36%(G + C)]的图谱是一致的。该区域占核DNA含量的45 - 50%,主要包含基因间序列和内含子序列。高(G + C)区域,即发现大多数编码序列的区域[38 - 68%(G + C)],其特征是有卫星带,表明存在转录惰性的串联重复序列家族。这些带可以相对高精度地分析卫星家族的丰度、碱基组成和序列差异。带型对每个物种都是独特的;即使是亲缘关系密切的物种也可以通过它们的碱基分布图谱轻易区分。通过流式细胞术测定,反刍动物核DNA含量的变化主要是由于这些重复序列家族丰度的变化。因此,发现驼鹿(A. alces)每个细胞含有8.85 ± 0.2 pg DNA,比反刍动物的平均含量多25%,而碱基分布曲线表明存在一种异常丰富的卫星序列,其(G + C)含量为52.6%。这种卫星的大小(1 kb)和序列与其他鹿科动物的卫星-I相对应,因此被命名为Alces-I。Alces-I的一个克隆重复序列长度为968 bp,(G + C)含量为52.6%,占核DNA的35%,即近300万个拷贝,比相关鹿科动物中该重复家族的平均阵列大小超出约300%。原位杂交表明该重复序列分布在所有62条近端着丝粒常染色体的着丝粒区域。Alces-I的GG、GA和AG数量远多于预期,而TA和CG双联体数量则少得多,这是所有串联重复序列的特征。该序列被判定与驯鹿(Rangifer tarandus)、狍(Capreolus capreolus)、黑麂(Muntiacus muntjac)、毛冠鹿(Muntiacus reevesi)以及叉角羚(Antilocapra americana),还有牛科动物牛(Bos taurus)和绵羊(Ovis aries)的卫星-I序列是直系同源的。基于形态特征提出的一个关于五种鹿科动物的初步系统树与之高度吻合。与共有序列的差异表明颠换比转换几乎多一倍,这表明替换是随机发生的,或者几乎是随机发生的。