摘要
目的:提取变形链球菌GS5葡糖基转移酶GTF(glucosyltransferase)催化活性区(CAT)的基因,利用IPTG诱导后获得可溶性表达,并检测活性。方法:利用PCR技术克隆CAT的基因片段,通过蛋白重组融合表达技术使其在大肠杆菌BL21中表达,利用异丙基硫代-β-D-半乳糖苷(IPTG)诱导含有质粒pET32-NusA-CAT的大肠杆菌,SDS-PAGE电泳进行检测;采用蒽酮硫酸法验证活性。结果:成功将CAT基因连入大肠杆菌BL21,获得可溶性的融合蛋白表达;并且融合蛋白具有生物学活性。结论:IPTG成功诱导克隆的变形链球菌葡糖基转移酶催化活性区基因并获得可溶性融合蛋白的表达,具有生物学活性。
Objective：To clone the catalytic region of Glucosyltransferase from Streptococcus mutans GS5 and express it in Escherichia coli BL21 by IPTG induction,obtain the soluble rGTF and activity assay. Method：PCR and construction of fusion protein technique,IPTG induction,anthrone-sulfuric acid assay. Result：The CAT gene was cloned correctly and its fusion protein was expressed in E.coli by IPTG induction. The SDS-PAGE results showed that the rGTF is soluble,and anthrone-sulfuric acid assay demonstrated its biological activity. Conclusion：The aimed gene and its fusion protein were gained successfully,and to be demonstrated its biological activity which provides a base to the further research.
出处
《中华老年口腔医学杂志》
2010年第2期94-97,共4页
Chinese Journal of Geriatric Dentistry
基金
国家自然科学基金资助项目(编号30440059)
