Arveiler B, Vincent A, Mandel J L
Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, Unité 184 de Biologie Moléculaire et de Génie Génétique de l'INSERM, Faculté de Médecine, Strasbourg, France.
Genomics. 1989 May;4(4):460-71. doi: 10.1016/0888-7543(89)90269-3.
We are using pulsed-field gel electrophoresis (PFGE) to establish a physical map of the human Xq28 region. We have identified a new probe 35.239 (DXYS64), localized in Xq28 by somatic hybrid mapping and belonging to a region of greater than 99% homology between the X and the Y chromosomes. PFGE data show that probes 35.239 and the polymorphic locus DXS115 (probe 767) map within a common 300-kb BssHII fragment. Both probes, in addition, hybridize to 575-kb BssHII and 590-kb ClaI fragments that contain the gene coding for coagulation factor VIII (F8C). The order F8C-DXS115-DXYS64 could be determined. Our results also provide evidence for linkage between the red/green color vision locus (RCP,GCP) and probes MD13 and T1.7 (GdX, DXS254) within a 750-kb ClaI fragment. Although the latter two probes are located within 50 kb of the 3' end of the G6PD gene, a G6PD cDNA probe did not hybridize to this fragment. G6PD, on the other hand, could be linked to F8C on a 290-kb BssHII fragment. All these data allow us to propose the order (RCP,GCP)-MD13-GdX-G6PD-F8C-DXS115-DXYS 64. We also linked probes St14 (DXS52), MN12 (DXS33), and DX13 (DXS15) to a member of a small family of X-linked dispersed sequences (DNF22S3) within a 575-kb BssHII fragment. The preliminary physical map presented here should be useful for further fine mapping of disease genes in the Xq28 region and should be helpful in orientating efforts toward the cloning of sequences close to the fragile X syndrome.
我们正在使用脉冲场凝胶电泳(PFGE)来构建人类Xq28区域的物理图谱。我们鉴定出了一个新探针35.239(DXYS64),通过体细胞杂交定位将其定位于Xq28,并且它属于X和Y染色体之间同源性大于99%的一个区域。PFGE数据表明,探针35.239和多态性位点DXS115(探针767)定位于一个共同的300kb BssHII片段内。此外,这两个探针均与包含凝血因子VIII(F8C)编码基因的575kb BssHII和590kb ClaI片段杂交。可以确定F8C-DXS115-DXYS64的顺序。我们的结果还为红/绿色觉位点(RCP、GCP)与750kb ClaI片段内的探针MD13和T1.7(GdX、DXS254)之间的连锁提供了证据。尽管后两个探针位于G6PD基因3'端的50kb范围内,但一个G6PD cDNA探针并未与该片段杂交。另一方面,G6PD可在一个290kb BssHII片段上与F8C连锁。所有这些数据使我们能够提出(RCP、GCP)-MD13-GdX-G6PD-F8C-DXS115-DXYS 64的顺序。我们还将探针St14(DXS52)、MN12(DXS33)和DX13(DXS15)与一个575kb BssHII片段内的X连锁分散序列小家族成员(DNF22S3)相连。此处呈现的初步物理图谱应有助于对Xq28区域的疾病基因进行进一步精细定位,并有助于指导针对克隆与脆性X综合征相关序列的工作。