利用爪蟾卵母细胞建立人类低密度脂蛋白受体基因表达系统
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国家自然科学基金(30070932)资助


Establishing the Expressive System of Human Low Density Lipoprotein Receptor Gene in the Oocyte of the Xenopus Laevis
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    摘要:

    目的 在爪蟾卵母细胞中建立人类低密度脂蛋白受体基因的表达系统,此系统可被用于降血脂和抗动脉粥样硬化的研究。方法和结果 冰块麻醉爪蟾,切取卵巢,获得V、VI期卵母细胞,经用1g/L胶原酶孵育过夜除去卵母细胞外层的卵巢膜和滤泡膜,培养3天后存活率仍可达95 %以上。用DiI标记的低密度脂蛋白荧光配体测定膜上结合荧光,发现V、VI期卵母细胞膜上未见有内源性低密度脂蛋白受体,如将人低密度脂蛋白受体的表达质粒p3.7低密度脂蛋白导入爪蟾卵母细胞核中,培养后用DiI标记的低密度脂蛋白配体测定可见膜上发出红色荧光,用抗低密度脂蛋白受体的抗体经免疫荧光检测法测定可见膜上有绿色荧光,免疫胶体金标记透射电镜检测可见膜上有内吞的胶体金颗粒。结论 外源性的人类低密度脂蛋白受体基因能在爪蟾V、VI期卵母细胞中表达,此低密度脂蛋白受体基因表达系统可用于中药降血脂抗动脉粥样硬化机理的研究。

    Abstract:

    Aim The expressive system of low density lipoprotein(LDL) receptor gene was established in the oocyte of the Xenopus laevis and the system is applied to investigation of lowering-lipids and anti-atheriosclerosis. Methods The fluorescence on the membrane with DiI-LDL were determined by culturing the oocytes in the period V and VI so as to judge whether the endogenous LDL receptor in oocyte of xenopus laevis existed. After then the p3.7LDL (an expression plasmid which contain human LDL receptor gene) was transmit to the nucleus of oocyte and the immunofluorescence and immunity colloid golden in the cell membrane were observed under microscope. Results The experiments proved there was no endogenous LDL receptor on the membrane of oocytes in the period V and VI. The exogenous LDL receptor can be expressed after the p3.7LDL was transmitted to the nucleus of oocyte. Conclusion Human LDL receptor gene can be expressed in Oocyte of Xenopus laevis and the system can be used in the investigation of lowering-lipids and anti-atheriosclerosis.

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沃兴德,范春雷,罗艳,钱颖,卢德赵,唐利华.利用爪蟾卵母细胞建立人类低密度脂蛋白受体基因表达系统[J].中国动脉硬化杂志,2005,13(2):138~142.

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  • 收稿日期:2004-05-08
  • 最后修改日期:2004-11-15
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