Weide R, Hontelez J, van Kammen A, Koornneef M, Zabel P
Department of Molecular Biology, Wageningen Agricultural University, Dreijenlaan, The Netherlands.
Mol Gen Genet. 1998 Aug;259(2):190-7. doi: 10.1007/s004380050804.
In order to isolate centromeric sequences from tomato (Lycopersicon esculentum) chromosome 6, a large-scale RAPD screen was performed. Among 1500 polymorphic RAPD markers tested, 100 were identified as chromosome 6-specific, using a L. esculentum substitution line carrying chromosome 6 from L. pennellii. Fifty-seven of these markers proved to originate from L. pennellii, and of these, 13 were genetically mapped between the morphological markers yellow virescent (yv) and thiaminless (tl), which flank the centromere. These markers could be assigned to three genetic loci, with 11 of the markers mapping to a single locus. Further resolution of this cluster was achieved using radiation-induced deletions that removed yv or tl but not the centromere. Seven markers were shown to be located outside all of the deletions. These seven markers and three of the other markers of the cluster were cloned and sequenced. Five of the clones are present as single-copy or low-copy-number sequences and five represent middle repetitive sequences. Three sequences show homology to the mammalian CENP-B binding box; clone AG12 contains two of these boxes and also shows homology to human satellite III.
为了从番茄(Lycopersicon esculentum)6号染色体中分离着丝粒序列,进行了大规模随机扩增多态性DNA(RAPD)筛选。在测试的1500个多态性RAPD标记中,利用携带来自潘那利番茄(L. pennellii)6号染色体的番茄(L. esculentum)代换系,鉴定出100个为6号染色体特异性标记。其中57个标记被证明源自潘那利番茄,在这些标记中,有13个在着丝粒两侧的形态标记黄化绿苗(yv)和无硫胺素(tl)之间进行了遗传定位。这些标记可被定位到三个基因位点,其中11个标记定位到单个位点。利用辐射诱导缺失进一步解析了这个簇,这些缺失去除了yv或tl,但没有去除着丝粒。七个标记被证明位于所有缺失区域之外。对这七个标记和该簇中的其他三个标记进行了克隆和测序。五个克隆以单拷贝或低拷贝数序列形式存在,五个代表中度重复序列。三个序列与哺乳动物着丝粒蛋白B(CENP - B)结合框具有同源性;克隆AG12包含其中两个框,并且还与人类卫星III具有同源性。