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肝脏特异性表达分泌型IgD转基因小鼠模型的构建

, PP. 286-294

Keywords: sIgD,肝脏特异性表达载体,HIDSMVK

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Abstract:

现有的遗传学研究认为高IgD综合征(HIDS)与甲羟戊酸激酶(MVK)基因发生突变有关,但具体的发病机制还不清楚,血清中IgD的升高与HIDS临床表征之间的关系还不知道.本研究运用融合PCR的方法构建肝脏特异性高表达嵌合分泌型IgD(csIgD)载体,并通过显微共注射,基因组PCR和Southernblotting鉴定出7只转基因F0代阳性小鼠.对F1代阳性小鼠在RNA和蛋白质水平分别进行了RT-PCR,Q-PCR和Westernblotting,ELISA检测.结果显示,转基因csIgD能够在肝脏特异性表达,并具有遗传稳定性.对阳性小鼠骨髓和脾脏细胞进行的流式分析发现,转基因csIgD的表达并未影响到B细胞的发育.形态解剖学观察,发现一些小鼠出现了皮肤破损和肝脾肾肿大的现象;病理切片显示,肝脾肾组织出现大量的细胞坏死和纤维样蛋白沉淀.TAIL-PCR(thermalasymmetricinterlacedPCR)检测结果表明,目的基因片段没有破坏位于小鼠5号染色体的MVK基因.本转基因小鼠模型的构建,有利于进一步对HIDS发病机制进行探讨.

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