CAJ | 학술논문

目的:将原核表达的多串重复人脑钠素蛋白裂解成单体.方法:利用高效表达质粒载体将串联5拷贝人脑钠素基因转入宿主菌E.coli M5219,以热诱导方式表达 5BNP融合蛋白,超声裂解收获 5BNP融合蛋白包涵体,制备性电泳分离纯化;凝血酶消化后,再经BMPS-Skatole 法裂解.结果:利用本文原核表达系统所表达的5BNP融合蛋白占菌体总蛋白的30%,采用制备性电泳可获得纯度＞95%的5BNP融合蛋白;凝血酶消化后,再经BNPS-Skatole法可以将5BNP融合蛋白裂解成BNP单体.结论:BMPS-Skatole法是裂解多串重复人脑钠素蛋白的良好方法.
목적:장원핵표체적다천중복인뇌납소단백렬해성단체.방법:이용고효표체질립재체장천련5고패인뇌납소기인전입숙주균E.coli M5219,이열유도방식표체 5BNP융합단백,초성렬해수획 5BNP융합단백포함체,제비성전영분리순화;응혈매소화후,재경BMPS-Skatole 법렬해.결과:이용본문원핵표체계통소표체적5BNP융합단백점균체총단백적30%,채용제비성전영가획득순도＞95%적5BNP융합단백;응혈매소화후,재경BNPS-Skatole법가이장5BNP융합단백렬해성BNP단체.결론:BMPS-Skatole법시렬해다천중복인뇌납소단백적량호방법.
Objactive:Tandem repeated human brain natriuretic peptide expressed in prokaryote was cleaved into BNP monomers. Methods: Five copies of BNP genes were tandem joined and inserted into the vector to construct highly expressed plasmid. The recombinant plasmids were transformed into the host cells E.coli. M5219, which expressed 5BNP recombinant proteins after temperature induction. Inclusion bodies containing fusion proteins of 5BNP were isolated by sonication. The fusion proteins were purified by preparative electrophoresis,and digested with thrombin and treated with BMPS-Skatole. Results: The fusion proteins of 5BNP were about 30% of the total proteins. The products purified by preparative electrophoresis, which were contained at least 95% of the fusion proteins of 5BNP, were digested with thrombin and cleaved into BNP monomers after treated with BMPS-Skatole. Conclusion: BMPS-Skatole was a good method for cleaving tandem repeated human brain natriuretic peptide.