Scheffer H, van der Lelie D, Aanstoot G H, Goor N, Nienhaus A J, van der Hout A H, Pearson P L, Buys C H
Hum Genet. 1986 Nov;74(3):249-55. doi: 10.1007/BF00282543.
From a human-Chinese hamster somatic cell hybrid a clone was derived containing chromosome 13 in duplicate as its only human material. This clone was used to construct a human chromosome 13-specific recombinant DNA-library. Overlapping Sau3AI DNA sequences (11.9-17.2 kb) from the cell hybrid were inserted into the lambda phage vector EMBL4. From eleven recombinants having a human insert thirteen putative unique DNA sequences were isolated and cloned into the plasmid vector pBR329. A human-mouse hybrid containing a human chromosome 13 with a deletion of 13q14 and lacking its undeleted homologue was constructed to be used in a selection procedure for DNA sequences belonging to band q14. Three probes originating from two different phages were assigned to 13q14 because they did not hybridise to DNA from this cell hybrid. One of these 13q14 probes detects a low frequency (2/44) MspI restriction fragment length polymorphism. The probes are now being used for screening a cosmid library to find adjacent polymorphic sequences with a RFLP information content suitable for application in the diagnosis of hereditary retinoblastoma.
从一个人-中国仓鼠体细胞杂种中获得了一个克隆，该克隆含有两份13号染色体，这是其唯一的人类物质。这个克隆被用来构建一个13号人类染色体特异性重组DNA文库。来自细胞杂种的重叠Sau3AI DNA序列（11.9 - 17.2 kb）被插入到λ噬菌体载体EMBL4中。从11个有人源插入片段的重组体中分离出13个假定的独特DNA序列，并克隆到质粒载体pBR329中。构建了一个含有13号人类染色体且13q14缺失且无未缺失同源物的人-鼠杂种，用于筛选属于q14带的DNA序列。来自两个不同噬菌体的三个探针被定位到13q14，因为它们不能与这个细胞杂种的DNA杂交。其中一个13q14探针检测到低频（2/44）MspI限制性片段长度多态性。这些探针现在正用于筛选黏粒文库，以寻找具有适合用于遗传性视网膜母细胞瘤诊断的RFLP信息含量的相邻多态性序列。
