与人精子抗原交叉的大肠埃希菌506株M_r36000蛋白的提取与纯化  

Extraction and purification of protein M r 36 000 from Escherichia coli 506 strain that cross reacts with human sperm antigens

作　　者：蒋玖[1] 陆德源[1] 郑佐娅[1] 

机构地区：[1]上海第二医科大学微生物学教研室

出　　处：《细胞与分子免疫学杂志》1998年第2期120-122,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：上海市高教局基金

摘　　要：以ＳＤＳ和超声波处理大量培养的Ｅ．ｃｏｌｉ５０６株制备全菌蛋白。首先采用（ＮＨ４）２ＳＯ４分段盐析对Ｍｒ３６０００蛋白进行初步纯化。发现与人精子交叉抗原的Ｍｒ３６０００蛋白位于６０％８０％和８０％９０％两个饱和硫酸铵盐析段中，并经与兔抗人精子抗体（ＲＡＨＳＡｂ）进行Ｗｅｓｔｅｒｎ印迹得以证实。将上述含Ｍｒ３６０００蛋白的盐析物合并，进行ＳｅｐｈａｄｅｘＧ７５和ＳｅｐｈａｄｅｘＧ２００柱层析，获得较纯的Ｍｒ３６０００蛋白，并对Ｍｒ３６０００纯化蛋白的特性进行了研究。用ＳＤＳＰＡＧＥ法测出纯化蛋白的Ｍｒ为３６０００；用Ｒｏｔｏｆｏｒ制备型等电聚焦电泳测定Ｍｒ３６０００蛋白的等电点为３８。The whole cell proteins were obtained by treatment of E coli 506 strain with sodium dodecyl sulfate( SDS) and ultrasonic wave A protein with molecular weight of 36 kd( M r36 000) was purified successively by ammonium sulfate prceipitation, column chromatography on Sephadex G 75 and Sephadex G 200 The purified protein could strongly react with rabbit anti human sperm antibody (RAHS Ab) on Western blotting Partial characterization of the protein including its relative molecular ( M r)and isoelectric point(PI)were investigated The M r was estimated to be about M r 36 000 as judged by SDS PAGE The PI of purified M r 36 000 protein was 3 8 determined by Rotofor preparative isoelectric focusing electrophoresis

关 键 词：大肠埃希菌 精子 共同抗原 纯化 抗原 

分 类 号：Q592.9[生物学—生物化学] R378.21[医药卫生—病原生物学]