一种简便高效制备伯氏疟原虫蛋白的实验方法
A simple and efficient methods to prepare soluable protein antigen from Plasmodium berghei
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摘要: 目的:探讨建立简便有效的大量制备鼠伯氏疟原虫蛋白的方法。方法:采集伯氏疟原虫感染晚期的大鼠外周血经肝素抗凝,通过白细胞滤器过滤去除白细胞,再使用30%阿拉伯胶溶液经密度梯度离心法分离含虫红细胞,经皂素溶血,收集纯化的原虫,虫体经超声波破碎后离心,上清液经SDS-PAGE电泳分析。采用蛋白凝胶灰度扫描方法分析蛋白电泳条带的分布。结果:从大鼠含虫外周血可分离制备出大量的可溶性疟原虫蛋白,对大鼠分离的疟原虫可溶性蛋白与小鼠来源的原虫可溶性蛋白比较,两种蛋白的电泳图谱完全相同。结论:利用大鼠模型可简便高效地制备大鼠伯氏疟原虫,结合超声波破碎法可提取足量的可溶性疟原虫蛋白抗原。Abstract: Objective:To establish a simple and convenient method of preparing and isolating soluble protein from Plasmodium berghei(P.berghei).Methods:SD rats were infected by mouse blood containing P.berghei,and then the rat peripheral blood was collected at the P.berghei-infected late stage.Furthermore the P.berghei-infected ＲBC were isolated by 25% arabic gum.The soluble protein was prepared from P.berghei by ultrasonication.The soluble protein was analyzed by SDS-PAGE,and compared with soluble protein from P.berghei-infected mice.Ｒesults:The large-scale yield of P.berghei can be prepared from P.berghei-infected rats,and the enough soluble protein can be obtained from P.berghei by ultrasonication.The proportion of each protein in soluble protein of P.berghei from infected rats is as the same as the protein from infected mice.Conclusions:The method of separating P.berghei from infected rats is a kind of simple and convenient way.The large-scale quantity of soluble protein antigen of P.berghei can be obtained by ultrasonication.
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