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耶尔森氏菌外膜蛋1(Yop1)的提取纯化与鉴定
Purificattion and Identification of Yersinia Outter-Membrane Protein 1
【摘要】 耶尔森氏菌外膜蛋白(Yops)在菌体中所占比例甚小,因而提取与纯化均有一定难度,国内尚无成功的先例。我们使用了一种盐析加电泳的方法提取了Yop1蛋白。实验结果表明,该法简便、易行且重复性好、回收率高。使用本法提取假结核耶氏菌外膜蛋白1(Yop1),自每升培基增殖的菌体中(约4.5g湿重),可获得纯品6.3mg,回收率高于国外同类研究的6倍以上[1]。一次性提取的Yop1蛋白在十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)中共显示出三条可辩认的蛋白带,其分子量依次为150KD,90~100KD及45~50KD。进一步的尿素-十二烷基磺酸钠-聚丙烯酰胺凝胶电泳结果表明,样品中的约100KD和50KD两种蛋白实际为150KD蛋白解聚后的二聚体和单体,根据这一结果,推测该种蛋白系三聚体,其单体分子量为45~50KD。
【Abstract】 ecause the proportion of Yersinia Outer membrane Proteins(Yops)is quite small in the bacteriumproteins,its purification is difficut and it had not been purified successfully in this country before,We purifiedthis protein by the method of salting out plus electrophoresis.The result showed that this method is simple,easy to handle,stable and with high recovery rate.The gain of pure protein 1 from 1 litre culture was 6.3mg.It Would show 3 visual bends on SDS PAGE gel and their molecular weight were 150,90~100 and 45~50KD respectively.It was deduced that the subunit molecular weight of this protein was 45~50 KD and the 150and 90~100 KD bends were represent their duplet and triplet forms respectively.
【Key words】 Yersinea; Yersinia Outer membrane Protein 1(Yop 1); Extrection; Purification; Identtifi-cation; Urea SDS PAGE; Molecular weight;
- 【文献出处】 地方病通报 ,ENDEMIC DISEASES BULLETIN (CHINA) , 编辑部邮箱 ,1994年01期
- 【分类号】R945
- 【被引频次】4
- 【下载频次】45