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目的:克隆和表达唾液链球菌尿素酶结构亚基UreA、UreB、UreC。方法:用已构建的含唾液链球菌57.I尿素酶基因簇的重组质粒为模板,分别克隆、双酶切、连接、转化,构建含结构基因的表达质粒,IPTG诱导表达,亲和层析纯化蛋白。结果:成功构建表达质粒,经诱导、亲和层析,获得高纯度、高浓度的UreA、UreB、UreC 3种目的蛋白。结论:成功诱导纯化唾液链球菌尿素酶的结构亚基,为今后各亚基的酶活分析和抗体制备等研究奠定基础。
OBJECTIVE: To clone and express urease structural subunits UreA, UreB, UreC of Streptococcus salivarius. Methods: The constructed recombinant plasmid containing Streptococcus salivarius 57.I urease gene cluster was cloned, double digested, ligated and transformed to construct the expression plasmid containing the structural gene, IPTG induced expression, affinity chromatography Purified protein. Results: The expression plasmids were successfully constructed and the target proteins of UreA, UreB and UreC with high purity and high concentration were obtained by induction and affinity chromatography. Conclusion: The structural subunits of purified urease of Streptococcus salivarius were successfully induced, which laid the foundation for future studies on enzyme analysis and antibody preparation of each subunit.