Morris D J, Adler I D, Robinson T J
Mammal Research Institute, University of Pretoria, South Africa.
Genomics. 1993 Feb;15(2):323-31. doi: 10.1006/geno.1993.1064.
Mouse x rat somatic cell hybrids were generated by fusing mouse cell lines that are heterozygous for reciprocal translocations involving the T42H and T9Ad breakpoints on mouse chromosome 11 (MMU11) to a thymidine kinase-negative (Tk-) rat cell line, RT2Tk-. Selection in HAT medium with geneticin disulfate (G418) resulted in some hybrid clones retaining only one derivative translocation chromosome with that part of MMU11 carrying the Tk-1 locus. Southern blot and PCR analyses of these hybrids were used to map the two breakpoints and 30 markers relative to them. The T42H breakpoint has been localized between Mpo and the Cola-1/Hox-2 cluster of loci and is proximal to the T9Ad breakpoint. The T9Ad breakpoint is proximal to the distal loci Tk-1, Gaa, D11Jkn1, and P4hb. The positions of 14 loci (Hox-2, Cola-1, Rara, Phb, Erba, Rnula-1, D11Pas1, Gfap, D11Mit13, D11Mit11, D11Mit12, Myla, Empb3 and Gh) have been further refined by their localization between the two breakpoints in band D. This study therefore improves the correlation of the genetic and physical maps of MMU11 and extends the known homology between MMU11 and human chromosome 17 (HSA17) by the assignment of three additional HSA17 markers, the profilin gene, Pfn, an anonymous marker, D17s28h, and the Crk oncogene, to above the T42H breakpoint; and the prohibitin gene, Phb, to between the T42H and T9Ad breakpoints in band D on MMU11.
通过将对涉及小鼠11号染色体(MMU11)上T42H和T9Ad断点的相互易位呈杂合状态的小鼠细胞系与胸苷激酶阴性(Tk-)大鼠细胞系RT2Tk-融合,产生了小鼠×大鼠体细胞杂种。在含有硫酸庆大霉素(G418)的HAT培养基中进行选择,结果一些杂种克隆仅保留了一条衍生易位染色体,该染色体上携带Tk-1位点的MMU11部分。对这些杂种进行Southern印迹和PCR分析,以绘制两个断点以及相对于它们的30个标记的图谱。T42H断点已定位在Mpo与Cola-1/Hox-2基因座簇之间,且位于T9Ad断点的近端。T9Ad断点位于远端基因座Tk-1、Gaa、D11Jkn1和P4hb的近端。通过将14个基因座(Hox-2、Cola-1、Rara、Phb、Erba、Rnula-1、D11Pas1、Gfap、D11Mit13、D11Mit11、D11Mit12、Myla、Empb3和Gh)定位在带D中的两个断点之间,其位置得到了进一步精确。因此,本研究提高了MMU11遗传图谱与物理图谱的相关性,并通过将另外三个HSA17标记(肌动蛋白结合蛋白基因Pfn、一个匿名标记D17s28h和Crk癌基因)定位到T42H断点上方;以及将抑制素基因Phb定位到MMU11带D中T42H和T9Ad断点之间,扩展了MMU11与人类17号染色体(HSA17)之间已知的同源性。