O'Connell P, Leach R J, Ledbetter D H, Cawthon R M, Culver M, Eldridge J R, Frej A K, Holm T R, Wolff E, Thayer M J
Howard Hughes Medical Institute, University of Utah Medical Center, Salt Lake City 84132.
Am J Hum Genet. 1989 Jan;44(1):51-7.
To better map the location of the von Recklinghausen neurofibromatosis (NF1) gene, we have characterized a somatic cell hybrid designated 7AE-11. This microcell-mediated, chromosome-transfer construct harbors a centromeric segment and a neo-marked segment from the distal long arm of human chromosome 17. We have identified 269 cosmid clones with human sequences from a 7AE-11 library and, using a panel of somatic cell hybrids with a total of six chromosome 17q breakpoints, have mapped 240 of these clones on chromosome 17q. The panel included a hybrid (NF13) carrying a der(22) chromosome that was isolated from an NF1 patient with a balanced translocation, t(17;22) (q11.2;q11.2). Fifty-three of the cosmids map into a region spanning the NF13 breakpoint, as defined by the two closest flanking breakpoints (17q11.2 and 17q11.2-q12). RFLP clones from a subset of these cosmids have been mapped by linkage analysis in normal reference families, to localize the NF1 gene more precisely and to enhance the potential for genetic diagnosis of this disorder. The cosmids in the NF1 region will be an important resource for testing DNA blots of large-fragment restriction-enzyme digests from NF1 patient cell lines, to detect rearrangements in patients' DNA and to identify the 17;22 NF1 translocation breakpoint.
为了更好地绘制冯·雷克林豪森神经纤维瘤病(NF1)基因的位置,我们对一个名为7AE - 11的体细胞杂种进行了特征分析。这个微细胞介导的染色体转移构建体包含一个着丝粒片段和来自人类17号染色体长臂远端的一个带有新霉素标记的片段。我们从一个7AE - 11文库中鉴定出了269个含有人类序列的黏粒克隆,并利用一组共有6个17号染色体q臂断点的体细胞杂种,将其中240个克隆定位到了17号染色体q臂上。该组包括一个携带der(22)染色体的杂种(NF13),它是从一名患有平衡易位t(17;22)(q11.2;q11.2)的NF1患者中分离出来的。53个黏粒定位到了一个跨越NF13断点的区域,该区域由两个最接近的侧翼断点(17q11.2和17q11.2 - q12)界定。通过在正常参考家系中进行连锁分析,对这些黏粒子集中的一部分RFLP克隆进行了定位,以更精确地定位NF1基因,并提高对这种疾病进行基因诊断的可能性。NF1区域的黏粒将成为检测NF1患者细胞系大片段限制性内切酶消化后的DNA印迹的重要资源,以检测患者DNA中的重排并确定17;22 NF1易位断点。