DESIGN OF HEPATITIS E VIRUS GENOTYPE 1 RECOMBINANT ORF3 PROTEIN BY CODON OPTIMIZATION METHOD

G. I. Alatortseva; Алаторцева Г. И.; A. V. Sidorov; Сидоров А. В.; L. N. Nesterenko; Нестеренко Л. Н.; L. N. Luhverchik; Лухверчик Л. Н.; M. V. Zhukina; Жукина М. В.; I. I. Amiantova; Амыантова И. И.; A. V. Milovanova; Мылованова А. В.; D. S. Vorobev; Воробьев Д. С.; Yu. I. Ammur; Аммур Ю. И.; M. I. Mikhailov; Михайлов М. И.; K. K. Kyuregyan; Кюрегян К. К.; V. S. Kichatova; Кичатова B. С.; I. A. Potemkin; Потемкин И. А.; O. V. Isaeva; Исаева О. В.; E. Yu. Malinnikova; Малинникова Е. Ю.; A. A. Karlsen; Карлсен А. А.; V. M. Blinov; Блинов В. М.; Z. Sh. Nurmatov; Нурматов З. Ш.; A. Z. Nurmatov; Нурматов А. З.; O. T. Kasymov; Касымов О. Т.; S. V. Zhavoronok; Жаворонок С. В.; V. V. Zverev; Зверев В. В.

doi:10.36233/0372-9311-2017-6-63-72

DESIGN OF HEPATITIS E VIRUS GENOTYPE 1 RECOMBINANT ORF3 PROTEIN BY CODON OPTIMIZATION METHOD

Authors: Alatortseva G.I.¹, Sidorov A.V.¹, Nesterenko L.N.¹, Luhverchik L.N.¹, Zhukina M.V.¹, Amiantova I.I.¹, Milovanova A.V.¹, Vorobev D.S.¹, Ammur Y.I.¹, Mikhailov M.I.², Kyuregyan K.K.², Kichatova V.S.², Potemkin I.A.², Isaeva O.V.², Malinnikova E.Y.², Karlsen A.A.², Blinov V.M.¹, Nurmatov Z.S.³, Nurmatov A.Z.³, Kasymov O.T.³, Zhavoronok S.V.⁴, Zverev V.V.¹
Affiliations:
1. Mechnikov Research Institute of Vaccines and Sera
2. Mechnikov Research Institute of Vaccines and Sera, Russian Medical Academy of Continuous Professional Education
3. Scientific Production Association «Preventive Medicine»
4. Belarusian State Medical University
Issue: Vol 94, No 6 (2017)
Pages: 63-72
Section: Articles
URL: https://microbiol.crie.ru/jour/article/view/232
DOI: https://doi.org/10.36233/0372-9311-2017-6-63-72
ID: 232

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Abstract

Aim. The development of the hepatitis E virus (HEV) genotype 1 full-size ORF3 recombinant polypeptide. Materials and methods. Escherichia coli strains, plasmid vectors, serological and clinical samples, ELISA reagent kits, molecular biological, bioinformatic, biotechnological, biochemical and serological methods. Results. HEV genotype 1 RNA had been isolated from clinical samples collected in Kyrgyzstan. DNA copy of subgenomic virus RNA had been cloned and used for further development of E.coli strains producing full-size recombinant protein ORF3 fused to E.coli beta-galactosidase. Codons optimization method was used in aim to increase expression level of recombinant protein. Recombinant protein ORF3 had been isolated from the inclusion bodies of the E.coli biomass and purified by size exclusion chromatography. Antigenic specificity of recombinant polypeptide had been confirmed by enzyme-linked immunosorbent assay and Western blotting with the specific sera. Conclusion. HEVgenotype 1 ORF3 recombinant antigen had been designed, and it’s applicability in diagnostic tests had been experimentally confirmed.

Keywords

hepatitis E virus, genotype 1 HEV, orf3 gene, recombinant ORF3 antigen, ELISA, Western blotting

References

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DESIGN OF HEPATITIS E VIRUS GENOTYPE 1 RECOMBINANT ORF3 PROTEIN BY CODON OPTIMIZATION METHOD

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About the authors

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