Brandes A, Thompson H, Dean C, Heslop-Harrison J S
Department of Cell Biology, John Innes Centre, Norwich, UK.
Chromosome Res. 1997 Jun;5(4):238-46. doi: 10.1023/a:1018415502795.
Nine repetitive DNA sequences, present in the haploid Arabidopsis thaliana genome in 7-300 copies, were hybridized in situ to metaphase and interphase chromosomes. Every sequence was detected on all five chromosome pairs, but was not evenly dispersed over the genome. Clusters of signals were found in particular regions of the centromeric heterochromatin, and each sequence showed a characteristic distribution pattern. Some sequences hybridized more strongly on different chromosomes, reflecting chromosome-specific amplification or the presence of homologous sequences. No hybridization signals could be detected on euchromatic regions. In situ hybridization on extended chromatin fibres showed that the pAL1 repeats are interrupted by another repetitive DNA sequence. A cosmid subclone (74A) contained a (GA)38 microsatellite motif, and hybridization with a (GA) oligonucleotide revealed that most of the hybridization sites of 74A correspond to the distribution of this microsatellite motif. The results show that the paracentromeric heterochromatin of A. thaliana chromosomes is composed not only of the tandemly arranged 180-bp repeat family pAL1/pAtMr, but also of some other repetitive sequences, thus giving a better understanding of the organization of sequences at the centromeres of A. thaliana.
9个重复DNA序列存在于单倍体拟南芥基因组中,拷贝数为7至300个,它们与中期和间期染色体进行原位杂交。每个序列在所有5对染色体上均被检测到,但在基因组中分布并不均匀。在着丝粒异染色质的特定区域发现了信号簇,并且每个序列都显示出特征性的分布模式。一些序列在不同染色体上杂交更强,这反映了染色体特异性扩增或同源序列的存在。在常染色质区域未检测到杂交信号。在伸展的染色质纤维上进行原位杂交表明,pAL1重复序列被另一个重复DNA序列打断。一个黏粒亚克隆(74A)包含一个(GA)38微卫星基序,与(GA)寡核苷酸杂交显示,74A的大多数杂交位点与该微卫星基序的分布相对应。结果表明,拟南芥染色体的近着丝粒异染色质不仅由串联排列的180 bp重复序列家族pAL1/pAtMr组成,还由一些其他重复序列组成,从而更好地了解了拟南芥着丝粒处序列的组织情况。