High-level Expression and Characterization of Fully Active Recombinant Conger Eel Galectins in Eschericia coli
High-level Expression and Characterization of Fully Active Recombinant Conger Eel Galectins in Eschericia coli
复制标题
全活性重组星鳗半乳糖凝集素在大肠杆菌中的高水平表达和表征
DOI：
10.1271/bbb.66.476
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
K. Muramoto
中科院分区：
文献类型：
--
作者：
T. Ogawa;C. Ishii;Y. Suda;H. Kamiya;K. Muramoto
利用pTV118N载体和大肠杆菌构建了重组鳗鱼Galectins的表达系统Congerins I和Congerins II。重组凝集素I和II可以以可溶性的活性形式获得，定量产率较高。位于Con I N-末端的Val和Leu密码子突变提高了表达效率。重组蛋白的纯化只需两个层析步骤即可完成。纯化的重组Congerins除了N-末端的乙酰基外，与天然的几乎相同，即它们具有相同的结构和糖类结合活性，表明N-末端的乙酰基对活性没有显著影响。
An expression system for recombinant conger eel galectins, congerins I and II, were constructed using the pTV 118N plasmid vector and Escherichia coli. Recombinant congerins I and II could be obtained in the soluble active form with high quantitative yield. Mutation of codons for Val and Leu located in the N-terminal region of Con I increased the expression efficiency. Purification of recombinant proteins were done by only two chromatographical steps from E. coli extract. The purified recombinant congerins were found to be almost the same as the native ones except for the acetyl group at the N-terminus; that is, they showed the same structures and carbohydrate binding activities, suggesting that N-terminal acetyl groups of congerins were not significant for activity.