摘要
SARS(严重急性呼吸综合征)是由一种新型的人类冠状病毒引起的。由于SARS冠状病毒3CL蛋白酶在病毒的复制翻译中起关键作用,所以成为抗SARS药物设计的关键靶标之一。该研究中,利用以pET28a为载体的高效原核表达系统,克隆表达出SARS冠状病毒3CL蛋白酶,然后由NTA-Ni2+亲和层析柱对表达的蛋白进行分离纯化。3CL蛋白酶在大肠杆菌表达系统中的成功表达为今后进一步筛选抗SARS病毒药物奠定基础。
A novel human coronavirus （SAR-CoV） has been reported to be the causative agent of severe a cute respiratory syndrome （SARS）. Sinee SARS-CoV 3C-like protease （3CL^pro） plays an important role in extensive proteolytic processing of viral replication, it is an attractive drug target for anti SARS agents. In this study, an efficient expression system, based on the pET28a vector, for producing SARS-3C-like proteinase, is presented. And the highly yielded expressed protease was purified by using NTA Ni^2＋ affinity chromatography. The results will be useful for the rational screening of anti-SARS drugs.
出处
《药物生物技术》
CAS
CSCD
2005年第4期219-223,共5页
Pharmaceutical Biotechnology
基金
This work was supported by anti-SARS drugs screening platform fund of the Minister of Science and Technology of China and anti-SARS drugs fund of China Pharmaceutical University(2003a03).
关键词
严重急性呼吸综合征
SARS冠状病毒
3CL蛋白酶
分子克隆
表达
分离纯化
Severe acute respiratory syndrome （SARS）, SARS coronavirus
3C-like proteinase, Molecular cloning, Expression, Purification
