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METHOD OF PRODUCTION OF RECOMBINANT ANTIGEN G2 OF HANTAVIRUS DOBRAVA IN CELLS OF E coli

机译:大肠杆菌细胞中汉布拉病毒汉布拉病毒重组抗原G2的生产方法

摘要

FIELD: biotechnology.SUBSTANCE: method is characterised in that the DNA of the structure RNAb indicated on Figure 1, which encodes the fused protein of three parts, where N-terminal position is green fluorescent protein GFP, central - peptide of 73 amino acid residues with the amino acid sequence of SRKKCNFATTPICEYDGNMVSGYKKVMATIDSFQAFNTSYIHYTDEQIEW KDPDGMLKDHLNILVTKDIDFDT, and C-terminal - light chain of double-stranded protein Kunitz-type inhibitor from potato tubers (PKPI-BI), are introduced into cells of E. coli. The cells transformed by this construction are cultured, the biomass is lysed, the insoluble fraction of the lysate is separated by centrifugation. The product of expression in the form of inclusion bodies is solubilised with the denaturant. Chromatography is carried out under denaturing conditions. The resulting product is used for detection of specific antibodies in serum of patients with hemorrhagic fever with renal syndrome.EFFECT: invention enables to obtain the recombinant antigen G2 of Hantavirus Dobrava with increased yield.6 dwg, 1 ex
机译:领域:生物技术。目的:该方法的特征在于图1所示的RNAb结构的DNA编码三部分的融合蛋白,其中N端是绿色荧光蛋白GFP,中心-73个氨基酸残基的肽将具有SRKKCNFATTPICEYDGNMVSGYKKVMATIDSFQAFNTSYIHYTDEQIEW KDPDGMLKDHLNILVTKDIDFDTDFDT氨基酸序列的马铃薯块茎(PKPI-BI)的双链蛋白Kunitz型抑制剂的C末端-轻链引入大肠杆菌细胞。培养通过该构建体转化的细胞,裂解生物质,通过离心分离裂解物的不溶部分。表达产物以包涵体的形式被变性剂溶解。色谱在变性条件下进行。所得产物用于检测肾综合征出血热患者血清中的特异性抗体。效果:本发明能够以增加的产率获得汉坦病毒杜夫拉娃的重组抗原G2。6dwg,1 ex

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