MAPPING HUMAN CHROMOSOME 11

一个完整的人类染色体图谱将大大有助于 了解人类生物学和分子疾病。 研究 本文提出的方法将产生人类染色体的限制性图谱 11并将该图谱与细胞遗传学和遗传图谱进行比对。 选择11号染色体是因为它编码基因， 与结构缺陷引起的疾病有关 致癌基因、生长因子、代谢紊乱、异常 发展，肿瘤，白血病，染色体重排和 脆弱的网站。 这项研究的另一个重要原因是， 大量的克隆探针已经被映射到这个 染色体 很明显，大量的订单 遗传标记对于完成完整的图谱是必不可少的。 而不是仅仅使用随机的DNA探针，我们将分离出一个独特的 人类11号染色体的一组有序DNA克隆，位于 在罕见的切割限制酶NotI的识别位点 和Sfil。 据估计，大约有300个克隆含有这些 将获得11号染色体上的识别位点，并将 足以产生完整的11号染色体限制性图谱 由50万到300万个碱基对的重叠DNA片段组成 在尺寸上。 探针将从一个中国人的图书馆中分离出来 仓鼠-人类细胞杂交，保留11号染色体作为其唯一的 染色体 将筛选阳性人类探针， 使用标记的寡核苷酸检测稀有切割位点的存在 NotI和SfiI序列。 这些序列将被定位在 11号染色体使用我们的快速定位细胞杂交面板， 将这条染色体分成大约20多个区域。 使用脉冲 场凝胶电泳和这些有序的标记，我们将 生成11号染色体的限制性图谱。 140个随机DNA 我们已经定位到11号染色体区域的标记， 我们目前分离的50多个随机探针 用于填充和完成有序的限制性酶切图谱。 那些 识别限制性片段多态性的探针将被 在连锁图谱中用于比对物理图谱和遗传图谱。 与疾病相关的基因和异常染色体断裂点 将位于大的限制性片段上， 在物理地图上分配它们。 此信息将 这对疾病的分子表征很重要。

A complete map for all human chromosomes would greatly facilitate understanding human biology and molecular disease. The studies proposed here will generate a restriction map of human chromosome 11 and align this map with the cytogenetic and genetic maps. Chromosome 11 was chosen since it encodes genes and has regions associated with diseases resulting from structural defects involved with oncogenes, growth factors, metabolic disorders, abnormal development, tumors, leukemias, chromosomal rearrangements and fragile sites. Another important reason for this study is the large number of cloned probes that have been mapped to this chromosome. It is clear that a very large number of ordered genetic markers are essential for accomplishing a complete map. Rather than employ only random DNA probes, we will isolate a unique set of ordered DNA clones for human chromosome 11 which are located at recognition sites for the rare cutting restriction enzymes NotI and SfiI. It is estimated that about 300 clones containing these recognition sites on chromosome 11 will be obtained, and will be sufficient to generate a complete chromosome 11 restriction map composed of overlapping DNA fragments 0.5 to 3 million base pairs in size. Probes will be isolated from a library of a Chinese hamster-human cell hybrid that retains chromosome 11 as its only chromosome. Positive human probes will be screened for the presence of rare cutter sites using labeled oligonucleotides for NotI and SfiI sequences. These sequences will be localized on chromosome 11 using our rapid mapping cell hybrid panel that will divide this chromosome into about tow dozen regions. Using pulsed field gel electrophoresis and these ordered markers, we will generate a restriction map for chromosome 11. The 140 random DNA markers that we have already mapped to regions of chromosome 11 and the more than 50 random probes we are currently isolating will be used to fill in and complete an ordered restriction map. Those probes recognizing restriction fragment polymorphisms will be employed in linkage maps to align the physical and genetic maps. Genes and abnormal chromosomal breakpoints associated with disease will be located on large restriction fragments which will precisely assign them on the physical map. This information will be significant for the molecular characterization of disease.