ПОЛУЧЕНИЕ РЕКОМБИНАНТНОГО АНАЛОГА КАПСИДНОГО БЕЛКА ВИРУСА ГЕПАТИТА Е 1 ГЕНОТИПА: КЛОНИРОВАНИЕ, ЭКСПРЕССИЯ, ОЧИСТКА, ОЦЕНКА АНТИГЕННЫХ СВОЙСТВ

Q3 Medicine
G. I. Alatortseva, Sidorov Av, L. Nesterenko, L. N. Luhverchik, V. Dotsenko, I. I. Amiantova, V. Y. Kabargina, A. V. Milovanova, D. S. Vorobev, Yu. I. Ammur, V. M. Blinov, A. Z. Nurmatov, Z. S. Nurmatov, D. A. Baiyzbekova, O. T. Kasymov, K. K. Kyuregyan, M. I. Mikhailov, S. V. Zhavoronok, Zverev Vv
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引用次数: 4

Abstract

Aim. The development of the hepatitis E virus (HEV) genotype 1 recombinant capsid protein. Materials and methods. Escherichia coli strains, plasmid vectors, serological and clinical samples, ELISA reagent kits, molecular biological, bioinformatic, biotechnological, biochemical and serological methods. Results. Using HEV genotype 1 DNA copy of subgenomic virus RNA we made E.coli strains producing recombinabt capsid protein, containing C-terminal fragment of ORF2 protein fused to E.coli beta-galactosidase. Recombinant protein ORF2 had been isolated from the inclusion bodies of the E.coli biomass and purified by size exclusion chromatography. By Western blotting it had been shown specific interaction of the recombinant polypeptide with anti-HEV IgG from pool of positive sera. Antigenic specificity of the recombinant polypeptide had been confirmed by enzyme-linked immunosorbent assay with sera of hepatitis E patients and reference groups: healthy donors, patients with hepatitis А, В, C, infectious mononucleosis and cytomegalovirus infection, HIV-infected patients. Conclusion. HEV genotype 1 ORF2 recombinant antigen had been developed, and its possible use in diagnostic tests had been experimentally shown.
复制型肝炎病毒e - 1型基因组蛋白:克隆、表达、精炼、抗原评估
的目标。戊型肝炎病毒(HEV)基因1型重组衣壳蛋白的研制。材料和方法。大肠杆菌菌株,质粒载体,血清学和临床样品,ELISA试剂盒,分子生物学,生物信息学,生物技术,生化和血清学方法。结果。利用亚基因组病毒RNA的HEV基因型1 DNA拷贝,我们制备了产生重组衣壳蛋白的大肠杆菌菌株,该蛋白含有ORF2蛋白的c端片段,与大肠杆菌β -半乳糖苷酶融合。从大肠杆菌生物量包涵体中分离得到重组蛋白ORF2,并采用大小排斥层析法纯化。Western blotting显示重组多肽与阳性血清抗hev IgG有特异性相互作用。重组多肽的抗原特异性已通过酶联免疫吸附试验在戊型肝炎患者和对照组血清中得到证实:健康献血者、А、В、丙型肝炎患者、传染性单核细胞增多症和巨细胞病毒感染患者、hiv感染者。结论。已经开发出HEV基因1型ORF2重组抗原,并通过实验证明其可能用于诊断试验。
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来源期刊
Zhurnal mikrobiologii, epidemiologii, i immunobiologii
Zhurnal mikrobiologii, epidemiologii, i immunobiologii Immunology and Microbiology-Immunology and Microbiology (miscellaneous)
CiteScore
1.40
自引率
0.00%
发文量
51
审稿时长
8 weeks
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