Vershinin A V, Schwarzacher T, Heslop-Harrison J S
John Innes Centre, Norwich, United Kingdom.
Plant Cell. 1995 Nov;7(11):1823-33. doi: 10.1105/tpc.7.11.1823.
Repetitive DNA sequences in the terminal heterochromatin of rye (Secale cereale) chromosomes have consequences for the structural and functional organization of chromosomes. The large-scale genomic organization of these regions was studied using the telomeric repeat from Arabidopsis and clones of three nonhomologous, tandemly repeated, subtelomeric DNA families with complex but contrasting higher order structural organizations. Polymerase chain reaction analysis with a single primer showed a fraction of the repeat units of one family organized in a "head-to-head" orientation. Such structures suggest evolution of chromosomes by chromatid-type breakage-fusion-bridge cycles. In situ hybridization and pulse field gel electrophoresis showed the order of the repeats and the heterogeneity in the lengths of individual arrays. After Xbal digestion and pulse field gel electrophoresis, the telomeric and two subtelomeric clones showed strong hybridization signals from 40 to 100 kb, with a maximum at 50 to 60 kb. We suggest that these fragments define a basic higher order structure and DNA loop domains of regions of rye chromosomes consisting of arrays of tandemly organized sequences.
黑麦(Secale cereale)染色体末端异染色质中的重复DNA序列对染色体的结构和功能组织有影响。利用拟南芥的端粒重复序列以及三个非同源、串联重复的亚端粒DNA家族的克隆,研究了这些区域的大规模基因组组织,这些家族具有复杂但截然不同的高级结构组织。用单一引物进行的聚合酶链反应分析表明,一个家族的部分重复单元以“头对头”方向排列。这种结构表明染色体通过染色单体型断裂-融合-桥循环进化。原位杂交和脉冲场凝胶电泳显示了重复序列的顺序以及各个阵列长度的异质性。经Xbal酶切和脉冲场凝胶电泳后,端粒克隆和两个亚端粒克隆在40至100 kb处显示出强烈的杂交信号,在50至60 kb处信号最强。我们认为,这些片段定义了由串联组织序列阵列组成的黑麦染色体区域的基本高级结构和DNA环结构域。