Genetic stability of the HA, NA, and NS genes of the recombinant vector virus FluNS1-124-Omp16 (H5N1) expressing the brucellar gene

Cover Page

Cite item

Full Text

Abstract

The recombinant strain Flu-NS1-124-Omp16 (H5N1) of the influenza virus expressing the brucellar Omp16 gene was constructed on the basis of the technology of reverse genetics for the purpose of developing vector antibrucellosis vaccine. The obtained recombinant strain is a genetically stable construction. This stability is confirmed by the comparative analysis of the nucleotide sequences of the HA, NA, and NS genes of the recombinant vector virus Flu-NS1-124-Omp16 (H5N1) expressing the Omp16 gene of the Brucella abortus (genBank: AAA59360.1). The comparative analysis showed that the nucleotide sequence of the Ns gene of the first and the fifth passage level of the Flu-NS1-124-Omp16 (H5N1) virus corresponded for 100% to the initial part of 12ААS2TC_124Omp16g containing the chimera NS1-124-Omp16 in the composition of DNA (deoxyribonucleic acid) plasmids pHW2000. Total identity with HA and NA genes of the strain А/AstanaRG/6:2/2009 (H5N1) was shown by the comparative analysis of the nucleotide sequences of HA and NA genes of the first and the fifth passage level of the recombinant strain Flu-NS1-124-Omp16 (H5N1). The recombinant vector virus Flu-NS1-124-Omp16 (H5N1) expressing the brucella Omp16 gene maintains the genetic stability during 5 passages in 10-day developing chicken embryos.

About the authors

S. O. Sadikaliyeva

Scientific-Research Institute of Biological Safety Problems

Author for correspondence.
Email: t.m.tlenchieva@mail.ru

Sandugash Sadikalieva, Junior research fellow

080409, Gvardeiskiy

Казахстан

K. T. Sultankulova

Scientific-Research Institute of Biological Safety Problems

Email: fake@neicon.ru
080409, Gvardeiskiy Казахстан

K. A. Shorayeva

Scientific-Research Institute of Biological Safety Problems

Email: fake@neicon.ru
080409, Gvardeiskiy Казахстан

V. M. Strochkov

Scientific-Research Institute of Biological Safety Problems

Email: fake@neicon.ru
080409, Gvardeiskiy Казахстан

O. V. Chervyakova

Scientific-Research Institute of Biological Safety Problems

Email: fake@neicon.ru
080409, Gvardeiskiy Казахстан

V. L. Zaitsev

Scientific-Research Institute of Biological Safety Problems

Email: fake@neicon.ru
080409, Gvardeiskiy Казахстан

K. K. Tabynov

Scientific-Research Institute of Biological Safety Problems

Email: fake@neicon.ru
080409, Gvardeiskiy Казахстан

N. T. Sandybayev

Scientific-Research Institute of Biological Safety Problems

Email: fake@neicon.ru
080409, Gvardeiskiy Казахстан

A. R. Sansyzbay

Scientific-Research Institute of Biological Safety Problems

Email: fake@neicon.ru
080409, Gvardeiskiy Казахстан

A. Yu. Egorov

HSC Development GmbH

Email: fake@neicon.ru
Tulln Австрия

References

  1. Racloz V., Schelling E., Chitnis N., Roth F., Zinsstag J. Persistence of brucellosis in pastoral systems. Rev. Sci. Tech. 2013; 32 (1): 61–70.
  2. Food and Agriculture Organization of the United Nations (FAO) (2010). Brucella melitensis in Eurasia and the Middle East. Proc. FAO technical meeting in collaboration with WHO and OIE, May 2009, Rome. FAO Animal Production and Health Proceedings No. 10. FAO, Rome. Available at: www.fao.org/docrep/012/i1402e/i1402e00.pdf ( Accessed on 5 April 2013).
  3. Egorov A., Brandt S., Sereinig S., Romanova J., Ferko B., Katinger D. et.al. Transfectant influenza A viruses with long deletions in the NS1 protein grow efficiently in Vero cells. J. Virol. 1998; 72(8): 6437–41.
  4. Fernandez-Sesma A. The influenza virus NS1 protein: inhibitor of innate and adaptive immunity. Infect. Disord. Drug Targets. 2007; 7(4): 336–43.
  5. Ferko B., Stasakova J., Romanova J., Kittel C., Sereinig S. Katinger H. et al. Immunogenicity and protection efficacy of replication-deficient influenza A viruses with altered NS1 genes. J. Virol. 2004; 78: 13037–45.
  6. Garcia-Sastre A., Biron C.A. Type 1 interferons and the virus-host relationship: a lesson in detente. Science. 2006; 312: 879–82.
  7. Строчков В.М., Червякова О.В., Султанкулова К.Т., Мамадалиев С.М., Зайцев В.Л., Шораева К.А. и др. Рекомбинантные штаммы А/AstanaRG/6:2/2009 и А/AstanaRG/5:3/2009 вируса гриппа, полученные методом обратной генетики. В кн.: Материалы международной конференции «Современное состояние генетики в Казахстане». Алматы; 2010; 63–5.
  8. Игнатьева А.В., Руднева И.А., Тимофеева Т.А., Шилов А. A., Забережный А.Д., Алипер Т.И. и др. Высокопродуктивный вирусреассортант, содержащий гемагглютинин и нейраминидазу пандемического вируса гриппа A/MOSCOW/01/2009(Н1N1). Вопросы вирусологии. 2011; 56 (4): 9–14.
  9. Гоцкина Т.М., Рыскельдинова Ш.Ж., Асанжанова Н.Н., Кыдырбаев Ж.К., Кожамкулов Е.М., Инкарбеков Д.И. и др. Возможность использования морских свинок как модельных животных для изучения иммунобиологических свойств рекомбинантных вирусов гриппа А, экспрессирующих бруцеллезные белки L7/L12 и Omp16. В кн.: Материалы международной научной конференции молодых ученых «Инновационное развитие науки в обеспечении биологической безопасности». Гвардейский; 2013; 47–54.
  10. De Wit E., Spronken M.I., Bestebroer T.M., Rimmelzwaan G.F., Osterhaus A.D., Fouchier R.A. Efficient generation and growth of influenza virus A/PR/8/34 from eight cDNA fragments. Virus Res. 2004; 103(1-2): 155–61.
  11. Hoffmann E., Neumann G., Kawaoka Y., Hobom G., Webster R.G. A DNA transfection system for generation of influenza A virus from eight plasmids. Proc. Natl. Acad. Sci. USA. 2010; 97(11): 6108–13.
  12. Ставицкий С.Б., Носков А.Н., Кравченко Т.Б. Молекулярная вакцина для профилактики бруцеллеза. Патент РФ № 2285538, 2006.
  13. WHO Guidance on development of influenza vaccine reference viruses by reverse genetics. Available at: whqlibdoc.who.int/hq/2005/WHO_CDS_CSR_GIP_2005.6.pdf.
  14. Leclercq S., Harms J.S., Oliveira S.C. Enhanced efficacy of DNA vaccines against an intracellular bacterial pathogen by genetic adjuvants. Curr. Pharm. Biotechnol. 2003; 4: 99–107.
  15. Pasquevich K.A., Estein S.M., Garcia Samartino C., Zwerdling A., Coria L.M., Barrionuevo P. et al. Immunization with recombinant Brucella spp. outer membrane proteins Omp16 or Omp19 in adjuvant induces specific CD41 and CD81 T cells as well as systemic and oral protection against Brucella abortus infection. Infect. Immun. 2008; 77(1): 436–45.
  16. Luo D., Ni B., Li P., Shi W., Zhang S., Han Y. et al. Protective immunity elicited by a divalentDNAvaccine encoding both the L7/L12 and Omp16 genes of Brucella abortus in BALB/c mice. Infect. Immun. 2006; 74: 2734–41.
  17. Wang X., Li M., Zheng H., Muster T., Palese P., Beg A.A. et al. Influenza A virus protein prevents activation of NFk-B and induction of alpha/bets interferon. J. Virol. 2000; 74(24): 11566–73.
  18. Kochs G., Martínez-Sobrido L., Lienenklaus S., Weiss S., GarcíaSastre A., Staeheli P. Strong interferon-inducing capacity of a highly virulent variant of influenza A virus strain PR8 with deletions in the NS1 gene. J. Gen. Virol. 2009; 90 (12): 2990–4.
  19. Liu M. Zhang Y., Liu C.G., Pan W.Q., Liu C.N., Yang T. Generation of High-yield vaccine strain wholly derived from Avian Influenza Viruses by Reverse Genetics. Chin. J. Biotech. 2006; 22: 720–6.
  20. Stech J., Garn H., Wegmann M., Wagner R., Klenk H.D. A new approach to an influenza live vaccine: modification of the cleavage site of hemagglutinin. Nat. Med. 2005; 11(6): 683–9.
  21. Hoffmann E., Webster R. G. Unidirectional RNA polymerase I-polymerase II transcription system for the generation of influenza A virus from eight plasmids. J. Gen. Virol. 2000; 81(12): 2843–7.

Supplementary files

Supplementary Files
Action
1. JATS XML
Download

Copyright (c) 2015 Sadikaliyeva S.O., Sultankulova K.T., Shorayeva K.A., Strochkov V.M., Chervyakova O.V., Zaitsev V.L., Tabynov K.K., Sandybayev N.T., Sansyzbay A.R., Egorov A.Y.

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).
Регистрационный номер и дата принятия решения о регистрации СМИ: серия ПИ № ФС77-77676 от 29.01.2020.


This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies