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

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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.

About the authors

G. I. Alatortseva

Mechnikov Research Institute of Vaccines and Sera

Author for correspondence.
Russian Federation

A. V. Sidorov

Mechnikov Research Institute of Vaccines and Sera

Russian Federation

L. N. Nesterenko

Mechnikov Research Institute of Vaccines and Sera

Russian Federation

L. N. Luhverchik

Mechnikov Research Institute of Vaccines and Sera

Russian Federation

M. V. Zhukina

Mechnikov Research Institute of Vaccines and Sera

Russian Federation

I. I. Amiantova

Mechnikov Research Institute of Vaccines and Sera

Russian Federation

A. V. Milovanova

Mechnikov Research Institute of Vaccines and Sera

Russian Federation

D. S. Vorobev

Mechnikov Research Institute of Vaccines and Sera

Russian Federation

Yu. I. Ammur

Mechnikov Research Institute of Vaccines and Sera

Russian Federation

M. I. Mikhailov

Mechnikov Research Institute of Vaccines and Sera, Russian Medical Academy of Continuous Professional Education

Russian Federation

K. K. Kyuregyan

Mechnikov Research Institute of Vaccines and Sera, Russian Medical Academy of Continuous Professional Education

Russian Federation

V. S. Kichatova

Mechnikov Research Institute of Vaccines and Sera, Russian Medical Academy of Continuous Professional Education

Russian Federation

I. A. Potemkin

Mechnikov Research Institute of Vaccines and Sera, Russian Medical Academy of Continuous Professional Education

Russian Federation

O. V. Isaeva

Mechnikov Research Institute of Vaccines and Sera, Russian Medical Academy of Continuous Professional Education

Russian Federation

E. Yu. Malinnikova

Mechnikov Research Institute of Vaccines and Sera, Russian Medical Academy of Continuous Professional Education

Russian Federation

A. A. Karlsen

Mechnikov Research Institute of Vaccines and Sera, Russian Medical Academy of Continuous Professional Education

Russian Federation

V. M. Blinov

Mechnikov Research Institute of Vaccines and Sera

Russian Federation

Z. Sh. Nurmatov

Scientific Production Association «Preventive Medicine»

Russian Federation

A. Z. Nurmatov

Scientific Production Association «Preventive Medicine»

Russian Federation

O. T. Kasymov

Scientific Production Association «Preventive Medicine»

Russian Federation

S. V. Zhavoronok

Belarusian State Medical University

Russian Federation

V. V. Zverev

Mechnikov Research Institute of Vaccines and Sera

Russian Federation

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Copyright (c) 2017 Alatortseva G.I., Sidorov A.V., Nesterenko L.N., Luhverchik L.N., Zhukina M.V., Amiantova I.I., Milovanova A.V., Vorobev D.S., Ammur Y.I., Mikhailov M.I., Kyuregyan K.K., Kichatova V.S., Potemkin I.A., Isaeva O.V., Malinnikova E.Y., Karlsen A.A., Blinov V.M., Nurmatov Z.S., Nurmatov A.Z., Kasymov O.T., Zhavoronok S.V., Zverev V.V.

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