【摘要】 目的从构建了人氨基末端脑利钠肽(aminoterm inal probrain natriuretic peptide,NTproBNP)基因的表达菌中抽取目的蛋白,并对其进行纯化、鉴定及稳定性的初步研究。方法取表达菌株BL21pGEX4T3NTproBNP,IPTG诱导表达,GSHagarose亲和纯化目的蛋白,提纯产物进行SDSPAGE电泳和Westernblot鉴定,并采用自动免疫分析仪检测其免疫反应性和稳定性。结果GSTNTproBNP融合蛋白表达水平高,可溶性好。SDSPAGE、免疫测定分析显示其相对分子量为34600,具有较高的特异免疫反应性和稳定性。结论NTproBNP能通过基因工程手段以GST融合蛋白的形式大量获取,在非变性条件下得到的GSTNTproBNP有较好的免疫反应活性和稳定性。更多还原

【Abstract】 Objective To extract human NT-proBNP from the E.coil which express GST-NT-proBNP fusion protein and identify the protein after purification,while its stability was studied.Methods BL21-pGEX-4T-3-NT-proBNP was amplified and induced by IPTG.The expression was purified by affinity chromatography through GSH-agarose and identified by SDS-PAGE and Western blot.The immunoreactivity and stability of the fusion protein was analyzed by commercial immune analyzer platform.Results The expression level of GST- NT-proBNP was high with good solubility.The fusion protein appeared in SDS-PAGE with molecular mass of 34 600 and could be recognized by NT-proBNP immune analyzer with good activity and stability.Conclusion Enough quantity of NT-proBNP could be produced in recombinant GST fusion protein form.Under non-denatural condition,GST-NT-proBNP with good immunoreactivity and stability could be obtained.更多还原