诱生型一氧化氮合成酶N端蛋白的诱导表达与纯化

Expression and Purification of Inducible Nitric Oxide Synthase

诱生型一氧化氮合成酶（ｉＮＯＳ）Ｎ端蛋白的表达，对阐明其功能和制备ｉＮＯＳ特异性抗体具有重要意义。本文对ｉＮＯＳ１１９６ＡＡ重组蛋白在大肠杆菌中的表达条件进行了优化，结果表明：在优化条件下，经ＩＰＴＧ诱导，重组蛋白表达量可达菌体蛋白的１５％２０％。为获得重组蛋白纯品，建立了Ｈｉｓ．ＢｉｎｄＴＭ柱亲和层析法和凝胶蛋白的两步纯化法。

Recombinant expression for sequence-specific domain of iNOS Nterminal is important for its functional elucidating and preparation of its specific antibody. In this report,optimiziation of expressive condition of iNOS(1196AA) expressible recombinant E.coli have been performed. The results showed that recombinant protein expressed at 15%￣20% in total host proteins after induction with optimized conditions. With the aim to obtain pure recombinant protein, we developed a method combined His.BindTM affinity chromotography with electrophoresis, and obtained fine purified (100% in pure) recombinant protein.