Gillett G T, McConville C M, Byrd P J, Stankovic T, Taylor A M, Hunt D M, West L F, Fox M F, Povey S, Benham F J
Galton Laboratory, University College London, United Kingdom.
Genomics. 1993 Feb;15(2):332-41. doi: 10.1006/geno.1993.1065.
High-dose irradiation hybrids containing fragments of chromosome 11 have been generated, with a view to isolating new region-specific markers. Forty-seven lines were scored for 34 markers: average retention was 6%. Fourteen lines contain markers from 11q14 to 11q23. One of these, Jo12, has 11q markers extending from tyrosinase (q14-q21) to PBGD (q23.3) plus one marker (TYRL, p11.2) from 11p. In situ hybridization using Alu PCR products from Jo12 as probe confirmed that the human DNA is derived from two regions, one in proximal 11p and a second, larger region in 11q23. Plasmid libraries of Alu PCR products from this and three other hybrids have been made. Six of eight recombinants identified as having single-copy inserts were mapped back to the regions of 11q22-q23 detected in the originating hybrid; only one mapped to a region not originally detected, and one was of hamster origin. These six clones provide new markers in 11q22-q23 that can be used directly for polymorphism studies. This series of hybrids is therefore a valuable resource for the rapid generation of markers from specific, defined regions of chromosome 11.
为了分离新的区域特异性标记,已构建了含有11号染色体片段的高剂量辐射杂种。对47个品系进行了34个标记的评分:平均保留率为6%。14个品系含有来自11q14至11q23的标记。其中一个品系Jo12,其11q标记从酪氨酸酶(q14-q21)延伸至PBGD(q23.3),外加一个来自11p的标记(TYRL,p11.2)。以Jo12的Alu PCR产物为探针进行原位杂交,证实人类DNA来自两个区域,一个在11p近端,另一个在11q23的较大区域。已构建了该杂种以及其他三个杂种的Alu PCR产物的质粒文库。鉴定为具有单拷贝插入片段的8个重组体中有6个被定位回原始杂种中检测到的11q22-q23区域;只有一个定位到一个最初未检测到的区域,还有一个是仓鼠来源。这六个克隆在11q22-q23中提供了可直接用于多态性研究的新标记。因此,这一系列杂种是从11号染色体特定、明确区域快速生成标记的宝贵资源。