β-伴大豆球蛋白β亚基的纯化及免疫活性鉴定  

Purification of β Subunit from β-conglycinin and Identification of Immunological Activity of the Prepared Subunit

作　　者：郑树贵[1,2] 曹松屹[3] 孙泽威[1] 秦贵信[1] 

机构地区：[1]吉林农业大学动物科技学院,吉林长春130118 [2]沈阳农业大学畜牧兽医学院,辽宁沈阳110161 [3]沈阳农业大学生物技术学院,辽宁沈阳110161

出　　处：《大豆科学》2009年第2期301-304,309,共5页Soybean Science

基　　金：国家自然科学基金重点资助项目(30430520)

摘　　要：等电点沉淀法和凝胶过滤层析提纯的β-伴大豆球蛋白经6mol·L-1脲变性后解聚为游离亚基,利用DEAE琼脂糖凝胶FF离子交换层析分离纯化β-伴大豆球蛋白亚基,并对纯化亚基进行透析复性。同时将β-伴大豆球蛋白免疫家兔制备抗血清,利用免疫印迹方法检测纯化的亚基与β-伴大豆球蛋白抗血清是否发生免疫反应。结果表明:利用DEAE琼脂糖凝胶FF阴离子交换层析可以纯化出高纯度的β-伴大豆球蛋白β亚基,纯化的β亚基可以与抗β-伴大豆球蛋白抗体结合,具有免疫活性。Purified β-conglycinin prepared by electric-point precipitation and filtration chromatography was denatured by 6 mol · L^- 1 urea and separated into free subunits. The subunits of β- conglycinin were isolated and purified by DEAE- Sepharose F F anion exchange chromatography and the prepared subunits were renatured by dialysis. The rabbits were immunized by β-conglycinin to produce β-conglycinin special antiserum. The immune responses between purified subunits and the antiserum were determined by immunoblotting. The results indicated ： pure β subunit could be prepared from β- conglycinin by DEAE-Sepharose F F anion exchange chromatography, and the purified β subunit could specially bind to β-conglycinin antiserum and this oroved that the subunit had immunological activity.

关 键 词：Β-伴大豆球蛋白 Β亚基 纯化 免疫活性 

分 类 号：S565.1[农业科学—作物学]