摘要
旨在通过优化AgrC_(TM6-7C)蛋白的纯化过程,获得高纯度的目的蛋白,为后续研究提供材料。详细探讨了金属离子螯合层析缓冲液中咪唑浓度、离子交换层析缓冲液的pH值对AgrC_(TM6-7C)蛋白纯化效果的影响,并研究了β-巯基乙醇对AgrC_(TM6-7C)蛋白聚集及其激酶活性的影响。经过优化后,在不影响AgrC_(TM6-7C)蛋白激酶活性的前提下,AgrC_(TM6-7C)蛋白在浓度和纯度上均有明显提高,纯度可达98%以上,产量可达15 mg/L。
The core objective of this paper is to obtain the protein of AgrC_（TM6-7C） with high purity by optimizing the purification processes,and provide the materials for further research.We investigated the influence of imidazole concentration in buffer used in the metal ion affinitychromatography and buffer with different pH used in the ion exchange chromatography on the purification of AgrC_（TM6-7C）.Moreover,we discussedthe effects of β-Mercaptoethanol on protein aggregation and kinase activity of AgrC_（TM6-7C）.The optimized purification method resulted in theobvious increase on the yield and purity of AgrC_（TM6-7C）,producing 15 mg with about 98% purity from one-liter culture medium.
出处
《生物技术通报》
CAS
CSCD
北大核心
2016年第7期59-65,共7页
Biotechnology Bulletin
基金
国家自然科学基金项目(21272031)
中央高校基本科研基金(DC201502020201)
