New types of mouse centromeric satellite DNAs
New types of mouse centromeric satellite DNAs
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DOI：
10.1007/s10577-005-2346-x
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发表时间：
2005-01-01
影响因子：
2.6
通讯作者：
Podgornaya, OI
中科院分区：
文献类型：
--
作者：
Kuznetsova, IS;Prusov, AN;Podgornaya, OI
基因组数据库不包含着丝粒区域的完整序列。我们创建了一个基于pUC 19的DNA片段库，从分离的间期核的染色中心。在这个文库中，我们发现了主要的卫星（MaSat）和两个新的卫星序列-MS 3和MS 4。对MS 3和MS 4序列进行计算机比对、片段弯曲状态分析和MAR模体搜索，并与小鼠大小卫星DNA（MiSat）进行比较，结果表明它们是新的卫星片段。MS 3和MS 4与总DNA经限制性内切酶酶切后的Southern杂交显示了卫星DNA的梯形特征。MS 3占总DNA的2.2%，其单体长度为150 bp。MS 4单体长300 bp，占总DNA的1.6%。在中期染色体上，MS 3和MS 4位于着丝粒区域。L929细胞周期中细胞核的FISH分析显示了MaSat、MiSat、MS 3和MS 4的相对位置。除了MaSat之外，所有映射的satDNA片段都属于G 0/G1期的染色中心的外层。MS 3可能参与了着丝粒的形成。小鼠基因组包含至少四种satDNA类型：富含AT（MaSat和MiSat）和富含CG（MS 3和MS 4）。
Genomic databases do not contain complete sequences of the centromeric regions. We created a pUC19-based library of DNA fragments from isolated chromocentres of interphase nuclei. In this library we have found major satellite (MaSat) and two new satellite sequences - MS3 and MS4. The computer analysis of MS3 and MS4 sequences by alignment, fragment curved state and search for MAR motifs in comparison with the mouse major and minor satellite (MiSat) DNA has shown them to be new satellite fragments. Southern blot of MS3 and MS4 with total DNA digested by restriction enzymes shows the ladder characteristic of satellite DNA. 2.2% of the total DNA consists of MS3, the monomer of which is 150 bp long. The MS4 monomer is 300 bp long and accounts for 1.6% of the total DNA. On metaphase chromosomes MS3 and MS4 are located at the centromeric region. FISH analysis of L929 nuclei during the cell cycle showed relative positions of MaSat, MiSat, MS3, and MS4. All mapped satDNA fragments except MaSat belong to the outer layer of the chromocentres in the G0/G1 phase. MS3 is likely to be involved in the centromere formation. The mouse genome contains at least four satDNA types: AT-rich (MaSat and MiSat), and CG-rich (MS3 and MS4).