摘要
目的:利用N端缺失10个氨基酸的葡激酶(recombinantstaphylokinase,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SDSPAGE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。
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.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2006年第5期58-62,共5页
China Biotechnology
关键词
重组葡激酶
诱导表达
蛋白纯化
Recombinant staphylokinase Inducation Purification
