【摘要】 采用匀浆粗提、硫酸铵分级沉淀、SephadexG-100、DEAE-32、SepharoseCL-6B等方法从水稻品种武育粳3号籽粒中分离纯化蔗糖合酶,所得的酶蛋白在非变性聚丙烯酰胺凝胶电泳胶上呈现一条深染色的蛋白酶带。HPLC测定其浓度为71.2%,SDS-PAGE测得的酶分子量为380ku。用得到的酶蛋白作为抗原,通过皮内多位点分次注射法免疫新西兰白兔获得免疫抗体,用此抗体成功地检测到水稻籽粒灌浆过程中蔗糖合酶蛋白的含量变化。更多还原

【Abstract】 Sucrose synthase (SuSy) was purified from rice grains (Oryza Sativa, L. . cv Wuyujing 3) using ammonium sulfate precipitation, gel filtration (SephadexG-100, Sepharose CL-6B) and ion exchange column chromatography (DEAE-32). The purity of SuSy protein identified by non-denatured PAGE was showed as one band in the gel. The molecular weight and the content of the purified SuSy protein were 3-8×102ku and 71.2% , respectively, determined by SDS-PAGE and HPLC. The antibody of SuSy was also prepared by immunizing rabbit with the purified SuSy protein as the antigen and using the antibody, we successfully detected the quantity of SuSy protein during rice grain-filling by western blotting.更多还原