Smith D I, Liu W, Ginzinger D, Green P, Smith S, Wang N D, Recchia F, Carolyn K, Drabkin H, Golembieski W
Department of Molecular Biology and Genetics, Wayne State University School of Medicine, Detroit, Michigan 48201.
Genomics. 1991 Sep;11(1):179-87. doi: 10.1016/0888-7543(91)90116-v.
A total of 5700 human chromosome 3-specific cosmid clones was isolated from a series of cosmid libraries constructed from somatic cell hybrids whose only human component was an entire chromosome 3 or a chromosome 3 containing an interstitial deletion removing 50% of long arm sequences. Several unique sequence chromosome 3-specific hybridization probes were isolated from each of 616 of these cosmids. These probes were then used to localize the cosmids by hybridization to a somatic cell hybrid deletion mapping panel capable of resolving chromosome 3 into nine distinct subregions. All 616 of the cosmids were localized to either the long or short arm of chromosome 3 and 63% of the short arm cosmids were more precisely localized. We have identified a total of 87 cosmids that contain fragments that are evolutionarily conserved. Fragments from these cosmids should prove useful in the identification of new chromosome 3-specific genes as well as in comparative mapping studies. The localized cosmids should provide excellent saturation of human chromosome 3 and facilitate the construction of physical and genetic linkage maps to identify various disease loci including Von Hippel Lindau disease and renal and small cell lung carcinoma.
从一系列由体细胞杂种构建的黏粒文库中分离出总计5700个特异于人类3号染色体的黏粒克隆,这些体细胞杂种中唯一的人类成分是一条完整的3号染色体或一条含有间质性缺失、去除了50%长臂序列的3号染色体。从这些黏粒中的616个中分别分离出几个特异于3号染色体的单拷贝序列杂交探针。然后通过与一个能够将3号染色体解析为9个不同亚区域的体细胞杂种缺失定位板杂交,利用这些探针来定位黏粒。所有616个黏粒都被定位到3号染色体的长臂或短臂上,并且63%的短臂黏粒被更精确地定位。我们总共鉴定出87个含有进化上保守片段的黏粒。来自这些黏粒的片段在鉴定新的3号染色体特异基因以及在比较图谱研究中应该会很有用。定位后的黏粒应该能很好地覆盖人类3号染色体,并有助于构建物理图谱和遗传连锁图谱,以鉴定包括冯·希佩尔-林道病、肾癌和小细胞肺癌在内的各种疾病位点。