Brown A, Copeland N G, Gilbert D J, Jenkins N A, Rossant J, Kothary R
Institut du cancer de Montréal, Centre de Recherche L.-C. Simard, P.Q., Canada.
Genomics. 1994 Apr;20(3):371-6. doi: 10.1006/geno.1994.1190.
We have previously identified a line of transgenic mice, Tg4, in which an hsp68-lacZ hybrid gene has inserted into the dystonia musculorum (dt) locus on chromosome 1. We have confirmed the localization of the Tg4 integration site to the proximal region of mouse chromosome 1 by interspecific backcross analysis. One end of the integration complex has been cloned and we have used single-copy probes from the flanking region to screen a mouse genomic library. Several overlapping lambda phage clones have been isolated and arranged into a contig spanning 75 kb of genomic DNA. Probes from the genomic contig have enabled us to characterize the wildtype and Tg4 loci. We report that the integration of the transgene was accompanied by a deletion of 45 kb of host genomic sequences with no other detectable rearrangement in the Tg4 genome.
我们之前鉴定出了一种转基因小鼠品系Tg4,其中一个hsp68-lacZ杂交基因插入到了1号染色体上的肌张力障碍小鼠(dt)基因座中。我们通过种间回交分析证实了Tg4整合位点定位于小鼠1号染色体的近端区域。整合复合体的一端已被克隆,并且我们使用侧翼区域的单拷贝探针筛选了小鼠基因组文库。已分离出几个重叠的λ噬菌体克隆,并将它们排列成一个跨越75 kb基因组DNA的重叠群。来自基因组重叠群的探针使我们能够对野生型和Tg4基因座进行表征。我们报告说,转基因的整合伴随着宿主基因组序列45 kb的缺失,在Tg4基因组中没有其他可检测到的重排。