Okazaki S, Tsuchida K, Maekawa H, Ishikawa H, Fujiwara H
Zoological Institute, Faculty of Science, University of Tokyo, Japan.
Mol Cell Biol. 1993 Mar;13(3):1424-32. doi: 10.1128/mcb.13.3.1424-1432.1993.
A pentanucleotide repetitive sequence, (TTAGG)n, has been isolated from a silkworm genomic library, using cross-hybridization with a (TTNGGG)5 sequence, which is conserved among most eukaryotic telomeres. Both fluorescent in situ hybridization and Bal 31 exonuclease experiments revealed major clusters of (TTAGG)n at the telomeres of all Bombyx chromosomes. To determine the evolutionary origin of this sequence, two types of telomeric sequence, (TTAGG)5 and a hexanucleotide repetitive sequence, (TTAGGG)4, which is conserved mainly among vertebrate and several invertebrate telomeres so far examined, were hybridized to DNAs from a wide variety of eukaryotic species under highly stringent hybridization conditions. The (TTAGGG)5 oligonucleotide hybridized to genomic DNAs from vertebrates and several nonvertebrate species, as has been reported so far, but not to any DNAs from insects. On the other hand, the Bombyx type of telomere sequence, (TTAGG)n, hybridized to DNAs from 8 of 11 orders of insect species tested but not to vertebrate DNAs, suggesting that this TTAGG repetitive sequence is conserved widely among insects.
利用与(TTNGGG)5序列的交叉杂交,从家蚕基因组文库中分离出一个五核苷酸重复序列(TTAGG)n,该序列在大多数真核生物端粒中是保守的。荧光原位杂交和Bal 31核酸外切酶实验均显示,(TTAGG)n主要聚集在家蚕所有染色体的端粒处。为了确定该序列的进化起源,在高度严格的杂交条件下,将两种类型的端粒序列,即(TTAGG)5和一种六核苷酸重复序列(TTAGGG)4(到目前为止,该序列主要在脊椎动物和几种已检测的无脊椎动物端粒中保守)与来自多种真核生物物种的DNA进行杂交。如先前报道,(TTAGGG)5寡核苷酸与脊椎动物和几种非脊椎动物物种的基因组DNA杂交,但不与任何昆虫的DNA杂交。另一方面,家蚕类型的端粒序列(TTAGG)n与所测试的11个昆虫目物种中的8个物种的DNA杂交,但不与脊椎动物的DNA杂交,这表明这种TTAGG重复序列在昆虫中广泛保守。