Francke U, Taggart R T
Proc Natl Acad Sci U S A. 1980 Jun;77(6):3595-9. doi: 10.1073/pnas.77.6.3595.
For comparative studies we have used the somatic cell hybridization approach to regionally map genes on the mouse X chromosome. Fibroblasts from a mouse with the balanced reciprocal translocation T(XD;16B5)16H were fused with a Chinese hamster cell line (V79/380-6) deficient in activity of the enzyme hypoxanthine phosphoribosyltransferase (HPRT). Interpecific cell hybrids were initially selected for retention of the mouse translocation chromosome carrying the Hprt gene. Subsequently, hybrid clones were counterselected to force segregation of this chromosome. Selected and counterselected hybrid clones were analyzed for their chromosome content by trypsin/Giemsa banding and for expression of the mouse forms of the X-linked enzymes HPRT and alpha-galactosidase (GALA) by isoelectric focusing. The results indicate that the breakpoint on the mouse X chromosome (in band XD) has separated the genes for HPRT (Hprt) and for GALA (Ags). Hprt is proximal to the breakpoint in region Xcen-XD and Ags is distal in region XD-Xter. The gene order in the mouse (centromere-Hprt-Ags) is therefore inverted when compared to the order of the homologous loci on the long arm of the human X (centromere-GALA-HPRT).
在比较研究中,我们采用体细胞杂交方法对小鼠X染色体上的基因进行区域定位。将携带平衡相互易位T(XD;16B5)16H的小鼠的成纤维细胞与次黄嘌呤磷酸核糖转移酶(HPRT)活性缺陷的中国仓鼠细胞系(V79/380 - 6)融合。种间细胞杂种最初被选择保留携带Hprt基因的小鼠易位染色体。随后,对杂种克隆进行反选择以促使该染色体发生分离。通过胰蛋白酶/吉姆萨染色分析选择和反选择的杂种克隆的染色体组成,并通过等电聚焦分析X连锁酶HPRT和α - 半乳糖苷酶(GALA)的小鼠形式的表达。结果表明,小鼠X染色体上的断点(在XD带)已将HPRT(Hprt)基因和GALA(Ags)基因分开。Hprt在Xcen - XD区域靠近断点,Ags在XD - Xter区域远离断点。因此,与人类X染色体长臂上同源基因座的顺序(着丝粒 - GALA - HPRT)相比,小鼠中的基因顺序(着丝粒 - Hprt - Ags)是颠倒的。