藻红蓝蛋白连接/异构酶重组体系  

Reconstitution system of phycoerythrocyanin lyase/isomerase

作　　者：周明[1] 王鲁[1] 郑雷[1] 赵开弘[1] 

机构地区：[1]华中科技大学环境科学与工程学院

出　　处：《华中科技大学学报（自然科学版）》2002年第8期114-116,共3页Journal of Huazhong University of Science and Technology(Natural Science Edition)

基　　金：国家自然科学基金资助项目 ( 39770 1 75 );湖北省科技攻关课题资助项目 ( 2 0 70 0 0 331 )

摘　　要：把层理鞭枝藻藻红蓝蛋白操纵子A ,E和F基因分别克隆在表达载体pET30中 ,转入大肠杆菌表达了相应蛋白质 .利用 pET30载体表达的蛋白质具有 6个连续组氨酸亲和标记 ,用Ni2 + 螯合亲和层析柱提纯了表达的蛋白质 .用这些纯化的蛋白质建立了藻红蓝蛋白α亚基体外重组系统 ,从而证明藻红蓝蛋白操纵子E和F基因编码层理鞭枝藻藻红蓝蛋白为连接Phycoerythrocyanin operon genes A, E, and F (pecA, pecE, and pecF) from Mastigocladus laminosus were inserted into expression vector pET30. After transforming E. coli, The coded proteins (HisTag-aPECA, HisTag-PECE, and HisTag-PECF) were overexpreesed. The overexpressed proteins were purified with Ni 2+ chelating affinity chromatography. By using the purified proteins, HisTag-PEC was enzymatically reconstituted with HisTag -PECA and PCB under catalysis of HisTag-PECE and HisTag-PECF. It was shown that pecE and pecF code the phycerythrocyanin lyase/isomerase.

关 键 词：藻红蓝蛋白 连接/异构酶 层理鞭枝藻 表达 重组 

分 类 号：S949.22[农业科学—水产养殖]