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Construction of a small Mus musculus repetitive DNA library: identification of a new satellite sequence in Mus musculus.

机译:小家鼠重复DNA文库的构建:小家鼠中新卫星序列的鉴定。

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摘要

We report the construction of a small library of recombinant plasmids containing Mus musculus repetitive DNA inserts. The repetitive cloned fraction was derived from denatured genomic DNA by reassociation to a Cot value at which repetitive, but not unique, sequences have reannealed followed by exhaustive S1 nuclease treatment to degrade single stranded DNA. Initial characterizations of this library by colony filter hybridizations have led to the identification of a previously undetected M. musculus minor satellite as well as to clones containing M. musculus major satellite sequences. This new satellite is repeated 10-20 times less than the major satellite in the M. musculus genome. It has a repeat length of 130 nucleotides compared with the M. musculus major satellite with a repeat length of 234 nucleotides. Sequence analysis of the minor satellite has shown that it has a 29 base pair region with extensive homology to one of the major satellite repeating subunits. We also show by in situ hybridization that this minor satellite sequence is located at the centromeres and possibly the arms of at least half the M musculus chromosomes. Sequences related to the minor satellite have been found in the DNA of a related Mus species, Mus spretus, and may represent the major satellite of that species.
机译:我们报告了含有小家鼠重复DNA插入片段的重组质粒小文库的构建。重复克隆的级分通过重新关联到Cot值从变性的基因组DNA衍生而来,在Cot值处重新退火但不是唯一的序列,然后进行彻底的S1核酸酶处理以降解单链DNA。通过集落滤器杂交对该库的初步表征导致鉴定了先前未检测到的小家鼠小卫星以及含有小家鼠小卫星序列的克隆。这种新卫星的重复量比小家鼠基因组中的主要卫星少10-20倍。与小家鼠主要卫星的重复长度为234个核苷酸相比,它的重复长度为130个核苷酸。小卫星的序列分析表明,它具有一个29个碱基对的区域,与主要卫星重复亚基之一具有广泛的同源性。我们还通过原位杂交表明,该次要卫星序列位于着丝粒,并且可能位于小家蝇M染色体的至少一半的臂上。在相关的Mus物种Ms spretus的DNA中发现了与小卫星有关的序列,可能代表该物种的主要卫星。

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