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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
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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
出处 《细胞与分子免疫学杂志》 CAS CSCD 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
关键词 大肠埃希菌 精子 共同抗原 纯化 抗原 E coli human sperm common antigen purification
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