Abrahams Brett S, Chong Annabelle C O, Nisha Maimun, Milette Danielle, Brewster David A, Berry Melissa L, Muratkhodjaev Farkhad, Mai Sabine, Rajcan-Separovic Evica, Simpson Elizabeth M
Graduate Program in Neuroscience, Department of Medical Genetics, University of Columbia, Vancouver, British Columbia, Canada.
Genesis. 2003 Jul;36(3):134-41. doi: 10.1002/gene.10205.
The evolving trend to use larger transgenes and their associated increased chance of unexpected genetic events mandates more careful characterization of transgenic mice. In characterizing our five new mouse strains transgenic for the BAC, bEMS4, we have identified the highest copy number reported to date: the stable incorporation of approximately 40 copies of a 194-kb expressed transgene in a single insertion site. We caution, however, that standard molecular techniques failed to identify a balanced translocation in another strain, and an inappropriate site of insertion in a third. Molecular cytogenetic analysis using metaphase FISH was the minimum level of characterization needed to reveal these unexpected genetic events. In addition, we combined FISH and SKY to identify the transgene at the breakpoints of the balanced translocation, t(3;9). This is the first description of a BAC-mediated chromosomal rearrangement and the first application of SKY to identify transgene-induced chromosomal rearrangements.
使用更大转基因及其相关的意外遗传事件发生几率增加的不断演变趋势，要求对转基因小鼠进行更仔细的表征。在对我们五个携带BAC、bEMS4的新小鼠品系进行表征时，我们发现了迄今为止报道的最高拷贝数：在单个插入位点稳定整合了约40个拷贝的194 kb表达转基因。然而，我们提醒，标准分子技术未能在另一个品系中鉴定出平衡易位，在第三个品系中也未能鉴定出不合适的插入位点。使用中期荧光原位杂交（FISH）的分子细胞遗传学分析是揭示这些意外遗传事件所需的最低表征水平。此外，我们将FISH和光谱核型分析（SKY）相结合，以在平衡易位t(3;9)的断点处鉴定转基因。这是对BAC介导的染色体重排的首次描述，也是SKY用于鉴定转基因诱导的染色体重排的首次应用。
