Allen M J, Jeffreys A J, Surani M A, Barton S, Norris M L, Collick A
Department of Genetics, University of Leicester, UK.
Nucleic Acids Res. 1994 Aug 11;22(15):2976-81. doi: 10.1093/nar/22.15.2976.
The human hypervariable minisatellite MS32 has a well characterised internal repeat unit array and high mutation rates have been observed at this locus. Analysis of MS32 mutants has shown that male germline mutations are polarised to one end of the array and frequently involve complex gene conversion-like events, suggesting that tandem repeat instability may be modulated by cis-acting sequences flanking the locus. In order to investigate the processes affecting MS32 mutation rate and mechanism, we have created transgenic mice harbouring an MS32 allele. Here we describe the organisation of eight transgenic insertions. Analysis of these transgenic loci by MVR-PCR and structural analysis of the junctions between mouse flanking DNA and the transgenic loci has shed light on mechanisms of integration and rearrangement of the tandem repeated transgenes. Sequence analysis of the mouse DNA flanking these transgenes has shown that 5 of the 8 insertions have integrated into mouse gamma satellite repeated sequence. This suggests a non-random integration of the MS32 transgene construct into the mouse genome.
人类高变微卫星MS32具有特征明确的内部重复单元阵列，并且在该位点观察到了高突变率。对MS32突变体的分析表明，雄性生殖系突变集中在阵列的一端，并且经常涉及复杂的基因转换样事件，这表明串联重复序列的不稳定性可能受到该位点侧翼顺式作用序列的调节。为了研究影响MS32突变率和机制的过程，我们创建了携带MS32等位基因的转基因小鼠。在此，我们描述了八个转基因插入片段的组织情况。通过MVR-PCR对这些转基因位点进行分析，并对小鼠侧翼DNA与转基因位点之间的连接处进行结构分析，揭示了串联重复转基因的整合和重排机制。对这些转基因侧翼的小鼠DNA进行序列分析表明，8个插入片段中有5个已整合到小鼠γ卫星重复序列中。这表明MS32转基因构建体在小鼠基因组中的整合是非随机的。
