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大豆的STR120卫星DNA:组织、进化及染色体特异性

The STR120 satellite DNA of soybean: organization, evolution and chromosomal specificity.

作者信息

Morgante M, Jurman I, Shi L, Zhu T, Keim P, Rafalski J A

机构信息

Du Pont Agricultural Products, Biotechnology Research, Experimental Station, Wilmington, DE 19880-0402, USA.

出版信息

Chromosome Res. 1997 Sep;5(6):363-73. doi: 10.1023/a:1018492208247.

Abstract

A highly repeated DNA sequence family, STR120, with tandemly arranged repetitive units (monomers) of approximately 120bp, has been identified in soybean [Glycine max (L.) Merr.]. Five related clones showing tandem repeats of a 120-bp-long monomer were isolated from a soybean genomic library. Results of Southern blotting experiments using three of the clones as probes onto genomic DNA digested with different restriction enzymes were in agreement with a tandem arrangement of these sequences in the genome. A total of 12 monomers were sequenced, showing considerable sequence heterogeneity. A consensus sequence of 126 bp was obtained that exhibits an average similarity of 81% to the sequenced units. In three of the clones identified, neighbouring units are significantly more similar to each other than to units from different clones; in the remaining two clones, however, similarity between the two units observed is low (70%), while the overall similarity between the two clones is high (95%). This indicates that in these cases the repetitive unit may be the dimer rather than the monomer. Based on the presence of direct repeats within each monomer, we suggest that the 120-bp monomer may itself have evolved by duplication of an ancestral 60-bp unit. The STR120 family distribution is limited to annual soybeans and is not found, at least at high-copy number, in related perennial soybeans or other members of the tribe Phaseolae. Fluorescence in situ hybridization (FISH) to metaphase chromosomes using four of the clones as probes shows that the number of chromosomal locations differs depending on the stringency conditions and goes from two to eight when the stringency is progressively lowered. The estimated copy number for one of the clones is from 5000 to 10000, but this may just represent a lower boundary for the whole family in consideration of the high sequence divergence observed within the family. FISH and sequence analysis therefore indicate that different subfamilies as well as higher-order repeat units are present in the STR120 family, very much like those in primate alpha satellite DNA, and that some of the subfamilies seem to exhibit divergence on a chromosomal basis.

摘要

在大豆[Glycine max (L.) Merr.]中鉴定出一个高度重复的DNA序列家族STR120,其重复单元(单体)串联排列,长度约为120bp。从大豆基因组文库中分离出五个相关克隆,显示出120bp长单体的串联重复。使用其中三个克隆作为探针,对用不同限制酶消化的基因组DNA进行Southern印迹实验的结果与这些序列在基因组中的串联排列一致。共对12个单体进行了测序,显示出相当大的序列异质性。获得了一个126bp的共有序列,其与测序单元的平均相似度为81%。在鉴定出的三个克隆中,相邻单元彼此之间的相似度明显高于与不同克隆中的单元;然而,在其余两个克隆中,观察到的两个单元之间的相似度较低(70%),而两个克隆之间的总体相似度较高(95%)。这表明在这些情况下,重复单元可能是二聚体而非单体。基于每个单体中存在直接重复序列,我们认为120bp的单体本身可能是由一个祖先60bp单元的复制进化而来。STR120家族的分布仅限于一年生大豆,至少在高拷贝数情况下,在相关的多年生大豆或菜豆族的其他成员中未发现。使用其中四个克隆作为探针,对中期染色体进行荧光原位杂交(FISH)表明,染色体定位的数量因严谨度条件而异,当严谨度逐渐降低时,从两个到八个不等。其中一个克隆的估计拷贝数为5000至10000,但考虑到该家族内观察到的高序列差异,这可能仅代表整个家族的下限。因此,FISH和序列分析表明,STR120家族中存在不同的亚家族以及高阶重复单元,这与灵长类动物α卫星DNA中的情况非常相似,并且一些亚家族似乎在染色体基础上表现出差异。

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