Kazama Yusuke, Sugiyama Ryuji, Matsunaga Sachihiro, Shibata Fukashi, Uchida Wakana, Hizume Masahiro, Kawano Shigeyuki
Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Chiba 277-8562, Japan.
J Plant Res. 2003 Aug;116(4):317-26. doi: 10.1007/s10265-003-0106-6. Epub 2003 Jul 8.
The major satellite DNAs of the dioecious plant Silene latifolia are represented by the repetitive sequences X43.1, RMY1 and members of the SacI family, which are located at the distal ends of chromosomes. To characterize the satellite DNAs at the distal ends of the chromosomes in S. latifolia ( Sl-distal-satDNA), we isolated a bacterial artificial chromosome clone (number 15B12) that contained multiple repeat sequences with KpnI restriction sites, and subcloned a portion of this sequence into a plasmid vector. Sequencing analysis confirmed that recognition or degenerate sites for KpnI were repeated 26 times at intervals of 310-324 bp in the inserted DNA. The phylogenetic tree that was constructed with the 26 KpnI repeat units contained clustered branches that were independent of the SacI family. It is clear that the KpnI repeat belongs to an Sl-distal-satDNA family that is distinct from the SacI family. We designated this family as " KpnI" after the restriction enzyme that does not have a site in the SacI family. Multi-colored fluorescent in situ hybridization was performed with the KpnI family and RMY1 probes under high stringency conditions. The results suggest that chromosome 7 is unique and that it carries the KpnI family at only one end.
雌雄异株植物宽叶蝇子草的主要卫星DNA由重复序列X43.1、RMY1和SacI家族成员代表，它们位于染色体的远端。为了表征宽叶蝇子草染色体远端的卫星DNA（Sl-远端-卫星DNA），我们分离了一个细菌人工染色体克隆（编号15B12），其包含多个带有KpnI限制性酶切位点的重复序列，并将该序列的一部分亚克隆到质粒载体中。测序分析证实，KpnI的识别或简并位点在插入DNA中以310 - 324 bp的间隔重复了26次。用26个KpnI重复单元构建的系统发育树包含与SacI家族无关的聚类分支。显然，KpnI重复序列属于一个与SacI家族不同的Sl-远端-卫星DNA家族。我们根据在SacI家族中没有位点的限制性酶将这个家族命名为“KpnI”。在高严谨条件下用KpnI家族和RMY1探针进行了多色荧光原位杂交。结果表明，7号染色体是独特的，并且它仅在一端携带KpnI家族。
