人血清中真核复制起始区DNA（Ors12DNA）结合蛋白的鉴定和纯化  

IDENTIFICATION AND PURIFICATION OF PROTEINS,WHICH BIND TO Ors12 DNA BEARING EUKARYOTIC ORIGIN OF DNA REPLICATION IN HUMAN SERUM

作　　者：王毅然[1] 朱祚铭[1] 陈家童[1] 

机构地区：[1]南开大学分子生物学研究所

出　　处：《南开大学学报（自然科学版）》1994年第2期8-15,共8页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：天津市21世纪青年科学基金

摘　　要：利用含有真核细胞复制起始区（ＯｒｉｇｉｎｏｆＤＮＡＲｅｐｌｉｃａｔｉｏｎ）的Ｏｒｓ１２ＤＮＡ作为探针，检测了人血清中与Ｏｒｓ１２ＤＮＡ特异结合的蛋白质。凝胶电泳迁移率改变实验表明人血清中存在特异Ｏｒｓ１２ＤＮＡ结合蛋白。实验还对影响Ｏｒｓ１２ＤＮＡ与蛋白质最适宜结合的各种因素进行了研究。苯酚处理反应体系及限制性内切酶的酶切位点保护实验都证实了Ｏｒｓ１２ＤＮＡ－蛋白质复合物的存在。利用硫酸铵盐析及ＤＮＡ亲和层析，对血清中的Ｏｒｓ１２ＤＮＡ结合蛋白进行了部分纯化，ＳＤＳ－聚丙烯酰胺凝胶电泳结果显示，Ｏｒｓ１２ＤＮＡ结合蛋白为一组非均一的蛋白质，亚基分子量分别为６４ｋＤ、４４ｋＤ和２４ｋＤ。Ors12 DNA, the eukaryotic origin of DZA replication, was used as a DNA probe to identify the specific Ors12 DNA binding proteins in human serum.Mobility shift assay showed that the apecific Ors12 DNA binding proteins exist in human serum.The factors influencing the binding of Ors12 DNA and serum proteins were studied.The Ors12 DNA-protein complex Was confirmed by treatment of DNA-Protein complet with phenol and assay of restriction cleaved sites protection. The Ors12 DNA binding proteins were partially purified by DNA affinity chromatography.SDS-PAGE shows that Ors12 DNA-binding proteins consist of several major components which have a molecular weight of 66 kD, 44kD and 24kD respectively.

关 键 词：0rs12DNA DNA结合蛋白 凝胶电泳迁移率 

分 类 号：Q513[生物学—生物化学]