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Archives of Virology

, Volume 164, Issue 10, pp 2505–2513 | Cite as

Identification of the optimal insertion site for expression of a foreign gene in an infectious hematopoietic necrosis virus vector

  • Jing-Zhuang Zhao
  • Li-Ming Xu
  • Miao Liu
  • Yong-Sheng Cao
  • Jia-Sheng Yin
  • Hong-Bai Liu
  • Tong-Yan LuEmail author
  • Zhen-Yu ZhangEmail author
Original Article

Abstract

Infectious hematopoietic necrosis virus (IHNV) was developed as a vector to aid the construction of vaccines against viral diseases such as viral hemorrhagic septicemia virus, spring viremia of carp virus, and influenza virus H1N1. However, the optimal site for foreign gene expression in the IHNV vector has not been determined. In the present study, five recombinant viruses with the green fluorescence protein (GFP) gene inserted into different genomic junction regions of the IHNV genomic sequence were generated using reverse genetics technology. Viral growth was severely delayed when the GFP gene was inserted into the intergenic region between the N and P genes. Real-time fluorescence quantitative PCR assays showed that the closer the GFP gene was inserted towards the 3ʹ end, the higher the GFP mRNA levels. Measurement of the GFP fluorescence intensity, which is the most direct method to determine the GFP protein expression level, showed that the highest GFP protein level was obtained when the gene was inserted into the intergenic region between the P and M genes. The results of this study suggest that the P and M gene junction region is the optimal site within the IHNV vector to express foreign genes, providing valuable information for the future development of live vector vaccines.

Notes

Acknowledgements

This study was supported by the Central Public-Interest Scientific Institution Basal Research Fund, CAFS (grant number HSY201804M and 2019ZD0704), the Natural Science Foundation of Heilongjiang Province (grant number QC2018039), the National Natural Science Foundation of China Grant (grant number 31802344), and the Heilongjiang Postdoctoral Program (LBH-Z18275).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest related to this work.

Supplementary material

705_2019_4366_MOESM1_ESM.doc (1.9 mb)
Supplementary material 1 (DOC 1905 kb)

References

  1. 1.
    Kurath G, Ahern KG, Pearson GD, Leong JC (1985) Molecular cloning of the six mRNA species of infectious hematopoietic necrosis virus, a fish rhabdovirus, and gene order determination by R-loop mapping. J Virol 53:469–476 (PubMed: 3838192) Google Scholar
  2. 2.
    Morzunov SP, Winton JR, Nichol ST (1995) The complete genome structure and phylogenetic relationship of infectious hematopoietic necrosis virus. Virus Res 38:175–192 (PubMed: 8578857) CrossRefGoogle Scholar
  3. 3.
    Kim KI, Cha SJ, Lee C, Baek H, Hwang SD, Cho MY, Jee BY, Park MA (2016) Genetic relatedness of infectious hematopoietic necrosis virus (IHNV) from cultured salmonids in Korea. Arch Virol 161:2305–2310 (PubMed: 27255747) CrossRefGoogle Scholar
  4. 4.
    Ammayappan A, LaPatra SE, Vakharia VN (2010) Molecular characterization of the virulent infectious hematopoietic necrosis virus (IHNV) strain 220-90. Virol J 7:10 (PubMed: 20085652) CrossRefGoogle Scholar
  5. 5.
    Adel M, Amiri AB, Dadar M, Breyta R, Kurath G, Laktarashi B, Ghajari A (2016) Phylogenetic relationships of Iranian infectious hematopoietic necrosis virus of rainbow trout (Oncorhynchus mykiss) based on the glycoprotein gene. Arch Virol 161:657–663 (PubMed: 26602428) CrossRefGoogle Scholar
  6. 6.
    Purcell MK, Marjara IS, Batts W, Kurath G, Hansen JD (2011) Transcriptome analysis of rainbow trout infected with high and low virulence strains of infectious hematopoietic necrosis virus. Fish Shellfish Immunol 30:84–93 (PubMed: 20883797) CrossRefGoogle Scholar
  7. 7.
    Enzmann PJ, Kurath G, Fichtner D, Bergmann SM (2005) Infectious hematopoietic necrosis virus: monophyletic origin of European isolates from North American genogroup M. Dis Aquat Organ 66:187–195 (PubMed: 16261933) CrossRefGoogle Scholar
  8. 8.
    Breyta R, Jones A, Stewart B, Brunson R, Thomas J, Kerwin J, Bertolini J, Mumford S, Patterson C, Kurath G (2013) Emergence of MD type infectious hematopoietic necrosis virus in Washington State coastal steelhead trout. Dis Aquat Organ 104:179–195 (PubMed: 23759556) CrossRefGoogle Scholar
  9. 9.
    Alonso M, Leong JC (2013) Licensed DNA vaccines against infectious hematopoietic necrosis virus (IHNV). Recent Pat DNA Gene Seq 7:62–65 (PubMed: 22670604) CrossRefGoogle Scholar
  10. 10.
    Ristow SS, LaPatra SE, Dixon R, Pedrow CR, Shewmaker WD, Park JW, Thorgaard GH (2000) Responses of cloned rainbow trout Oncorhynchus mykiss to an attenuated strain of infectious hematopoietic necrosis virus. Dis Aquat Organ 42:163–172 (PubMed: 11104067) CrossRefGoogle Scholar
  11. 11.
    Larragoite ET, Tacchi L, LaPatra SE, Salinas I (2016) An attenuated virus vaccine appears safe to the central nervous system of rainbow trout (Oncorhynchus mykiss) after intranasal delivery. Fish Shellfish Immunol 49:351–354 (PubMed: 26772477) CrossRefGoogle Scholar
  12. 12.
    Romero A, Figueras A, Thoulouze MI, Bremont M, Novoa B (2008) Recombinant infectious hematopoietic necrosis viruses induce protection for rainbow trout Oncorhynchus mykiss. Dis Aquat Organ 80:123–135 (PubMed: 18717065) CrossRefGoogle Scholar
  13. 13.
    Rouxel RN, Tafalla C, Merour E, Leal E, Biacchesi S, Bremont M (2016) Attenuated infectious hematopoietic necrosis virus with rearranged gene order as potential vaccine. J Virol 90:10857–10866 (PubMed: 27681130) CrossRefGoogle Scholar
  14. 14.
    Emmenegger EJ, Biacchesi S, Mérour E, Glenn JA, Palmer AD, Brémont M, Kurath G (2018) Virulence of a chimeric recombinant infectious haematopoietic necrosis virus expressing the spring viraemia of carp virus glycoprotein in salmonid and cyprinid fish. J Fish Dis 41:67–78 (PubMed: 28799647) CrossRefGoogle Scholar
  15. 15.
    Romero A, Figueras A, Tafalla C, Thoulouze MI, Bremont M, Novoa B (2005) Histological, serological and virulence studies on rainbow trout experimentally infected with recombinant infectious hematopoietic necrosis viruses. Dis Aquat Organ 68:17–28 (PubMed: 16465830) CrossRefGoogle Scholar
  16. 16.
    Rouxel RN, Mérour E, Biacchesi S, Brémont M (2016) Complete protection against influenza virus H1N1 strain A/PR/8/34 challenge in mice immunized with non-adjuvanted Novirhabdovirus vaccines. Plos One 11:e0164245 (PubMed: 27711176) CrossRefGoogle Scholar
  17. 17.
    Zhao W, Spatz S, Zhang Z, Wen G, Garcia M, Zsak L, Yu Q (2014) Newcastle disease virus (NDV) recombinants expressing infectious laryngotracheitis virus (ILTV) glycoproteins gB and gD protect chickens against ILTV and NDV challenges. J Virol 88:8397–8406 (PubMed: 24829337)l CrossRefGoogle Scholar
  18. 18.
    Zhao JZ, Xu LM, Zhang ZY, Liu M, Cao YS, Yin JS, Liu HB, Lu TY (2019) Recovery of recombinant infectious hematopoietic necrosis virus strain Sn1203 using the mammalian cell line BHK-21. J Virol Methods 265:84–90 (PubMed:30615899) CrossRefGoogle Scholar
  19. 19.
    Zhao JZ, Xu LM, Liu M, Zhang ZY, Yin JS, Liu HB, Lu TY (2017) Autophagy induced by infectious hematopoietic necrosis virus inhibits intracellular viral replication and extracellular viral yields in epithelioma papulosum cyprini cell line. Dev Comp Immunol 77:88–94 (PubMed: 28760360) CrossRefGoogle Scholar
  20. 20.
    Zhao JZ, Xu LM, Liu M, Cao YS, LaPatra SE, Yin JS, Lu TY (2017) Preliminary study of an oral vaccine against infectious hematopoietic necrosis virus using improved yeast surface display technology. Mol Immunol 85:196–204 (PubMed: 28285182) CrossRefGoogle Scholar
  21. 21.
    Novoa B, Romero A, Mulero V, Rodríguez I, Fernández I, Figueras A (2006) Zebrafish (Danio rerio) as a model for the study of vaccination against viral haemorrhagic septicemia virus (VHSV). Vaccine 24:5806–5816 (PubMed: 16777275) CrossRefGoogle Scholar
  22. 22.
    Nzonza A, Lecollinet S, Chat S, Lowenski S, Merour E, Biacchesi S, Bremont M (2014) A recombinant Novirhabdovirus presenting at the surface the E glycoprotein from West Nile Virus (WNV) is immunogenic and provides partial protection against lethal WNV challenge in BALB/c mice. Plos One 9:e91766 (PubMed: 24663075) CrossRefGoogle Scholar
  23. 23.
    Zhang Z, Zhao W, Li D, Yang J, Zsak L, Yu Q (2015) Development of a Newcastle disease virus vector expressing a foreign gene through an internal ribosomal entry site provides direct proof for a sequential transcription mechanism. J Gen Virol 96:2028–2035 (PubMed: 25872740) CrossRefGoogle Scholar
  24. 24.
    Carnero E, Li W, Borderia AV, Moltedo B, Moran T, Garcia-Sastre A (2009) Optimization of human immunodeficiency virus gag expression by newcastle disease virus vectors for the induction of potent immune responses. J Virol 83:584–597 (PubMed: 19004953) CrossRefGoogle Scholar
  25. 25.
    Wertz GW, Moudy R, Ball LA (2002) Adding genes to the RNA genome of vesicular stomatitis virus: positional effects on stability of expression. J Virol 76:7642–7650 (PubMed: 12097578) CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Jing-Zhuang Zhao
    • 1
  • Li-Ming Xu
    • 1
  • Miao Liu
    • 1
  • Yong-Sheng Cao
    • 1
  • Jia-Sheng Yin
    • 1
  • Hong-Bai Liu
    • 1
  • Tong-Yan Lu
    • 1
    Email author
  • Zhen-Yu Zhang
    • 2
    Email author
  1. 1.Heilongjiang River Fishery Research Institute Chinese Academy of Fishery SciencesHarbinPeople’s Republic of China
  2. 2.State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbinChina

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