SINES AND LINES CLUSTER IN DISTINCT DNA FRAGMENTS OF GIEMSA BAND SIZE
SINES AND LINES CLUSTER IN DISTINCT DNA FRAGMENTS OF GIEMSA BAND SIZE
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DOI：
10.1007/bf00292382
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发表时间：
1989-11-01
期刊：
影响因子：
1.6
通讯作者：
MANUELIDIS, L
中科院分区：
文献类型：
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作者：
CHEN, TL;MANUELIDIS, L
通过原位杂交，以Alu重复为代表的短穿插重复DNA元件(Sine)主要位于Giemsa-light人类中期染色体带上。相反，L1长散布重复序列(线)家族优先聚集在Giemsa-Dark带中。这些正弦/线图案还通常对应于早期和后期复制带图案。为了在结构上可见的染色体带和散布重复序列的框架之间提供分子联系，我们使用脉冲场凝胶电泳(PFGE)研究了正弦和线形杂交的模式。散布的正弦和品系与0.2-3兆碱基对(Mb)的特定大小片段进行高强度杂交。使用适当的限制性内切酶和脉冲场条件，可以描绘出许多富含正弦或线条的片段，并且是相互排斥的。以与主系家族相关的人内源性逆转录病毒重复序列进行的对照研究，描绘了0.07-0.4Mb的片段子集，强度不同。因此，这些数量较少的重复序列似乎也选择性地聚集在比染色体环(60-120kb)更大的DNA结构域中。总而言之，PFGE研究独立地证实了相邻DNA环上散布的重复序列的聚集。重复基序的选择性聚集可能有助于大染色体结构域的特殊结构或功能特性，如染色质延伸/凝聚或复制特性。在某些情况下，由这些重复序列定义的DNA片段接近串联卫星阵列的大小。
By in situ hybridization, short interspersed repeated DNA elements (SINEs), exemplified by Alu repeats, are located principally in Giemsa-light human metaphase chromosome bands. In contrast, the L1 family of long interspersed repeats (LINEs) preferentially cluster in Giemsa-dark bands. These SINE/LINE patterns also generally correspond to early and later replication band patterns. In order to provide a molecular link between structurally visible chromosome bands and a framework of interspersed repeats, we investigated patterns of SINE and LINE hybridization using pulse-field gel electrophoresis (PFGE). Interspersed SINEs and LINEs hybridize with high intensity to specific size fragments of 0.2-3 megabase pairs (Mb). Using appropriate restriction enzymes and pulse-field conditions a number of fragments were delineated that were either SINE or LINE rich, and were mutually exclusive. Control studies with a human endogenous retroviral repeat that is related in sequence to the major LINE family, delineated a subset of fragments of 0.07-0.4 Mb with unequal intensity. Thus these less numerous repeats also appear to cluster selectively in DNA domains that are larger than a chromosome loop (60-120 kb). In summary, PFGE studies independently confirm the clustering of interspersed repeats on contiguous DNA loops. Selective clustering of repeat motifs may contribute to special structural or functional properties of large chromosome domains, such as chromatin extension/condensation or replication characteristics. In some cases the DNA fragments defined by these repeats approach the size of tandem satellite arrays.