摘要
目的为进一步研究肿瘤的治疗提供高质量的可溶性多亮氨酸重复区免疫球蛋白样蛋白(sLRIG1)。方法用重组sLRIG1的杆状病毒(Bac-sLRIG1)感染sf9昆虫细胞,表达的蛋白经提纯,获得sLRIG1蛋白,用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹进行分析鉴定。结果使用杆状病毒表达载体系统(BEVS)成功表达了重组sLRIG1蛋白,最佳感染复率为5,孵育时间为72 h,经鉴定蛋白表达正确,纯度高达95%。结论用BEVS可成功获得大量高纯度重组sLRIG1蛋白,可满足后续实验的需要。
Objective To provide high quality soluble leucine-rich repeats and immunoglobulin-like domains 1 (sLRIG1) protein for further research on the treatment of malignant tumors. Methods Sf9 insect cells were infected with the recombinant sLRIG1 Baculovirus encoding the extra-cellular domain of human LRIG1 (gene No. AF381545). The supernatant of the culture medium was collected and the recombinant sLRIG1 was purified by the Ni Sepharose Fast Flow Column and the DEAE Sepharose Fast Flow Column. The obtained protein was identified by SDS-PAGE technique and Western blotting. Results The sLRIG1 protein was successfully expressed and its expression reached peak at 72 hours after infection when the multiplicity of infection was 5. The purity was over 95%. Conclusion The sLRIG1 protein was successfully preparated via Baculovirus expression vector system. The purified protein had a high quality and was suitable for the further research on the treatment of malignant tumors.
出处
《中国临床神经外科杂志》
2012年第8期476-478,共3页
Chinese Journal of Clinical Neurosurgery
基金
教育部博士点新教师基金No.200804861039
国家自然科学基金No.30973073
关键词
可溶性LRIG1
杆状病毒蛋白表达系统
蛋白表达
纯化
鉴定
Soluble leucine-rich repeats and immunoglobulin-like domains 1 (sLRIG1)
Baculovirns Expression Vector System (BEVS)
Preparation
Identification