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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 coliM5 2 19,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
Objective: The prokaryotic expression of multiple strings of repeated human brain natriuretic protein fragmentation monomer. Methods: The 5-copy human brain natriuretic peptide gene was transferred into E. coli host cell E. coli M5 2 19 by high efficiency expression plasmid vector. 5BNP fusion protein was expressed by heat induction. Incubation of 5BNP fusion protein was obtained by ultrasonic lysis and purified by preparative electrophoresis. After enzymatic digestion, it is cleaved by BMPS-Skatole method. Results: The 5BNP fusion protein expressed by the prokaryotic expression system accounted for 30% of the total bacterial proteins, and the purity of> 95% of the 5BNP fusion protein was obtained by preparative electrophoresis. After digestion with thrombin and BNPS-Skatole method, 5BNP fusion protein to BNP monomer. CONCLUSIONS: The BMPS-Skatole method is a good way to cleave multiple strings of repeated human brain natriuretic peptides