摘要
目的:对蛋白四聚体(Tetramer)进行改造,简化其制备过程,提高制备效率,节约成本。方法:利用基因工程技术,将肽-HLA-A2和BirA酶的底物肽(BSP)形成一个融合蛋白,该融合蛋白在原核中以包涵体的形式被表达,对表达的包涵体蛋白进行稀释复性、酶切和纯化后,再在BirA酶的作用下对其生物素化,最后将其连接到荧光素标记的亲和素上,即形成了改造的Tetramer。结果:获得了可溶性的蛋白四聚体。结论:该方法制备过程简单,蛋白复兴效率高,成本低,值得继续深入研究。
Objective To simplify the preparation procedure,raise the preparation efficiency and reduce the cost,the tetramer was improved by technology of genetic engineering.Methods:The gene of a fusion protein was constructed by linking the sequence of the peptide-HLA-A2 and the substrate of BirA enzyme. Then this gene was expressed in BL21(DE3) and the product protein was in a form of inclusion body.The inclusion body protein was refolded by dilution method and biotinylated at the role of BirA enzyme.At last,this protein was linked with fluorescein-labeled avidin and a improved tetramer was formed.Results: An advanced tetramer was produced.Conclusion This new method is convenient,high efficient and has a low cost.It is worth studying thoroughly in the future.
出处
《山西农业大学学报（社会科学版）》
2007年第5期11-15,共5页
Journal of Shanxi Agricultural University:Social Science Edition
