Gajović Srećko, Mitrecić Dinko, Augustincić Lana, Iaconcig Alessandra, Muro Andrés F
Croatian Institute for Brain Research, School of Medicine, Univeristy of Zagreb, Croatia.
Transgenic Res. 2006 Apr;15(2):255-9. doi: 10.1007/s11248-006-0003-6.
Uniform genetic background of inbred mouse strains is essential in experiments with genetically modified mice. In order to assess Add2 (beta-adducin) function, its null mutation was produced in embryonic stem cells derived from 129Sv mouse and the subsequently obtained mouse mutants were backcrossed for 6 generations with C57BL/6JOlaHsd strain. Comparison of brain proteins between mutated and control animals by two-dimensional gels linked to mass spectroscopy analysis showed expression of Snca (alpha-synuclein) in the mutated animals, but unexpectedly not in the control C57BL/6JOlaHsd mice. Comparison between C57BL/6JOlaHsd and C57BL/6NCrl mice confirmed the presence of a deletion encompassing Snca and in addition Mmrn1 (multimerin1) loci in C57BL/6JOlaHsd strain. The segregation of mutated Add2 together with an adjacent part of the chromosome 6 derived from 129Sv mice, rescued the loss of these two genes in knockout mice on C57BL/6JOlaHsd background. The fact that Add2 knockout was compared with the C57BL/6JOlaHsd mouse strain, which is actually a double knockout of Snca and Mmrn1 emphasizes a need for information provided by commercial suppliers and of exact denominations of substrains used in research.
近交系小鼠品系的统一遗传背景在转基因小鼠实验中至关重要。为了评估Add2（β-内收蛋白）的功能，在源自129Sv小鼠的胚胎干细胞中产生了其无效突变，随后获得的小鼠突变体与C57BL/6JOlaHsd品系回交6代。通过与质谱分析相关联的二维凝胶对突变动物和对照动物的脑蛋白进行比较，结果显示Snca（α-突触核蛋白）在突变动物中表达，但在对照C57BL/6JOlaHsd小鼠中却意外未表达。C57BL/6JOlaHsd和C57BL/6NCrl小鼠之间的比较证实了C57BL/6JOlaHsd品系中存在一个包含Snca以及Mmrn1（多聚体蛋白1）基因座的缺失。源自129Sv小鼠的突变Add2与6号染色体的相邻部分一起分离，挽救了C57BL/6JOlaHsd背景敲除小鼠中这两个基因的缺失。将Add2敲除与实际上是Snca和Mmrn1双敲除的C57BL/6JOlaHsd小鼠品系进行比较这一事实，强调了商业供应商提供信息以及研究中使用的亚系确切名称的必要性。
