水稻NADP-苹果酸酶的原核表达、纯化和抗体制备  

Prokaryotic Expression,Purification and Antibody Production of Rice (Oryza sativa L.)NADP-Malic Enzyme

作　　者：程玉祥[1] 柳参奎[1] 

机构地区：[1]东北林业大学盐碱地生物资源环境研究中心,哈尔滨150040

出　　处：《植物生理学通讯》2007年第5期847-851,共5页Plant Physiology Communications

基　　金：国家高科技研究发展计划(2002AA241111);东北林业大学校立科研基金

摘　　要：构建了水稻NADP-ME_2基因cDNA的原核表达载体pQE30,并诱导表达出有生物学功能的融合蛋白。用Ni-NTA琼脂糖亲和层析纯化出NADP-ME_2融合蛋白,并测定了融合蛋白酶学特性(V_(max)、K_m、K_(cat)、底物特异性)。用纯化的NADP- ME_2融合蛋白免疫家兔,制备出抗水稻NADP-ME特异性抗体。全蛋白双向电泳后Western印迹表明水稻中至少有4个NADP-ME家族蛋白质成员。Rice NADP-ME2 cDNA was constructed into prokaryotic expression vector （pQE30）, and its bio- logically active fusion protein was expressed. NADP-ME2 fusion protein was purified by Ni-NTA agarose affinity chromatography, and its enzyme properties such as Vmax, Km,Kcat, and substrate specificity were detected. Purified NADP-ME2 fusion protein was used to immunolize rabbit. Anti-rice NADP-ME specific antibody was obtained. Four members of NADP-ME family protein had been detected by Western blot from rice total proteins after two-dimensional gel electrophoresis.

关 键 词：水稻 NADP-ME2 原核表达 酶学特性 抗体 

分 类 号：Q943.2[生物学—植物学]