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目的对FVB/N品系Mxi1基因敲除(knock out,KO)(Mxi1~(~(-/-)))小鼠进行遗传背景转换,为研究疾病发病机制提供更多种类的基因修饰小鼠。方法将FVB/N品系杂合子小鼠(Mxi1~(+/-))与异性C57BL/6品系野生型(wild type,WT)小鼠(Mxi1~(+/+))杂交,获得杂合子F1代后将其与C57BL/6品系Mxi1+/+小鼠回交,再选择F2杂合子与C57BL/6品系Mxi1~(+/+)小鼠回交,依此再回交8次后,子代互交。利用鼠尾DNA进行基因型鉴定及微卫星DNA(msDNA)检测,组织信使RNA(mRNA)和组织蛋白检测背景转换效率。结果基因型鉴定为Mxi1~(-/-)的互交子代小鼠msDNA检测结果与WT型C57BL/6小鼠一致,与FVB/N小鼠不同。PCR和Western印迹结果显示,随机选取的任意自交子代纯合子小鼠体内不表达Mxi1的mRNA和蛋白质,成功获得C57BL/6品系Mxi1~(-/-)小鼠。结论通过杂交-回交的方式成功获得的新品系Mxi1~(-/-)小鼠能稳定地将突变基因传递给子代。这一成果为疾病动物模型的建立提供了更多的选择。
Objective To investigate the genetic background of Mxi1 knock out (KO) mice (Mxi1 ~ (~ (- / -))) in FVB / N strain and provide more kinds of genetically modified mice for studying the pathogenesis of the disease. Methods Hybrid FVB / N mice (Mxi1 ~ (+/-)) and heterozygous C57BL / 6 wild type (WT) mice (Mxi1 ~ (+ / + After that, it was backcrossed with Mxi1 + / + mice of C57BL / 6 strain, and backflora of F2 heterozygotes was backcrossed with Mxi1 ~ (+ / +) mice of C57BL / 6 strain. Mutual exchange. Genomic identification and microsatellite DNA (msDNA) detection were performed using murine tail DNA, and background conversion efficiency was measured using tissue messenger RNA (mRNA) and tissue protein. Results The results of msDNA analysis of Mxi1 ~ (- / -) crossbred offspring mice were consistent with that of WT C57BL / 6 mice, which was different from that of FVB / N mice. PCR and Western blotting results showed that MXI1 ~ (- / -) mice were successfully obtained without any expression of Mxi1 mRNA and protein in any randomly selected inbred homozygous mice. Conclusion The new strain of Mxi1 ~ (- / -) mice, successfully obtained by hybridization-backcross, can stably transmit the mutant gene to progenies. This result provides more choices for the establishment of disease animal models.