摘要
大豆种子经过浸提、离心、硫酸铵逐级沉淀和D-半乳糖-Epoxy-Sepharose-4B亲和层析,获得单一的大豆凝集素(Soybean agglutinin SBA)样品。纯化的大豆凝集素经pH8.9的聚丙烯酰胺凝胶电泳(胶浓度7.5%),可以显示一条清晰的着色带。在血凝活性鉴定中,采用2%兔细胞悬液,测定凝集红细胞的能力和检测它们与糖结合后对红细胞的凝集作用的影响。结果表明,使兔红细胞50%凝集的蛋白质最低浓度为4μg/mL左右。浓度为0.1mg/ml凝集素溶液分别与D-半乳糖、D-氨基半乳糖、异丙基--βD-硫代半乳糖苷(IPTG)、а—乳糖、几丁质溶液结合后,使兔红细胞50%凝集的糖配体溶液浓度分别为1mg/ml、2.5mg/ml、10mg/ml3、0 mg/ml,而几丁质几乎对SBA没有影响。
Soybean agglutinin （SBA） was purified from the excellent seeds of Soybean by extraction, fraction with （NH4）2SO4, D-galactose-Epoxy-Sepharose-4B affinity chromatography. The purified SBA showed single PAGE outline and its ability to agglutination for erythrocytes of rabbits. During the research of the comparison of the ability of heamagglutination after combining 5 kinds of sugar ligands, D- galactose, D-galactosamine, IPTG, a-lactose and chitin were added in turn into the solution of SBA to observe the inhibition of the ability to agglutination for erythrocytes of rabbits. The result showed that the ability of SBA to agglutination for erythrocytes was inhibited, and the lowest concentration of the five kinds of sugar ligids to make 50% erythrocytes agglutinated was in turn 1mg/mL,2.5mg/mL,10mg/mL,30 mg/ mL, but chitin had no effect to inhibition of the ability to agglutination for erythrocytes of rabbits.
出处
《家畜生态学报》
2006年第6期57-60,共4页
Journal of Domestic Animal Ecology
基金
国家自然科学基金项目(编号:A200340)
关键词
大豆凝集素
纯化
血凝
糖配体
Soybean agglutination
Purification
Heamagglutination
Sugar ligand
