重组葡激酶工程菌的构建方法

目的：利用N端缺失10个氨基酸的葡激酶（recombinant staphylokinase，rSaK）重组质粒，构建了表达可溶性rSaK126蛋白的工程菌，并研究不同条件下工程菌诱导表达目的蛋白含量的差异及纯化途径。 方法：采用细菌活化和培养方法诱导目的蛋白，并用SDS-PAGE测其含量，应用层析技术纯化蛋白。 结果：成功构建表达重组葡激酶的工程菌，表达的重组葡激酶蛋白约占菌体总蛋白的50％，经纯化后回收率为60%，纯度达99%以上。结论：成功构建高效表达重组葡激酶的工程菌，并获得了高含量、高纯度的目的蛋白。

Inducement and purification of recombinant staphylokinase

Objective: Using the recombinant staphylokinase there is a lack of ten amino acids in the N end to construct engineering expressing soluble protein. Research the content of recombinant staphylokinase gained at different induced conditions and the way of purification. Methods: The aim protein of recombinant staphylokinase induced using the activation and cultivation of bacteria and its content was measured with SDS-PAGE. The protein was purified with chromatography. Result: The content of induced recombinant staphylokinase was about 50% of total protein. After purification, the rate of recollected aim protein was 60% and its purity was over 99%. Conclusion: The engineering of induced aim protein of recombinant staphylokinase were successfully constructed. The high expression and purified protein was acquired.