Advertisement

Archives of Microbiology

, Volume 199, Issue 2, pp 357–364 | Cite as

Identification and functional analysis of the GTPV bidirectional promoter region

  • Hui Zhang
  • Zhihua Sun
  • Na Zhang
  • Zhiqiang Li
  • Pengyan Wang
  • Qiang Fu
  • Yan Ren
  • Xuehua Shao
  • Yu Zhang
  • Zhiru Guo
  • Chuangfu ChenEmail author
Original Paper

Abstract

The goat pox chick embryo-attenuated virus (GTPV) has been developed as an effective vaccine that can elicit protective immune responses. It possesses a large genome and a robust ability to express exogenous genes. Thus, this virus is an ideal vector for recombinant live vaccines for infectious diseases in ruminant animals. In this study, we identified a novel bidirectional promoter region of GTPV through screening named PbVV(±). PbVV(±) is located between ETF-l and VITF-3, which are transcribed in opposite directions. A new recombinant goat pox virus (rGTPV) was constructed, in which duplicate PbVV(+) was used as a promoter element to enhance Brucella OMP31 expression, and duplicate PbVV(−) was used as a promoter element to regulate enhanced green fluorescent protein (EGFP) at the same time as the selection marker. PbVV(−) promoter activity was compared to that of the P7.5 promoter of vaccinia virus, as measured by EGFP expression; the fluorescence intensity of EGFP expressed in cells was confirmed by fluorescence microscopy and flow cytometry. PbVV(+) promoter activity was measured by Brucella OMP31 expression. Interaction with the anti-Brucella-OMP31 monoclonal antibody was confirmed by western blotting, and OMP31 mRNA expression was assessed by qRT-PCR. The results of this study will be useful for the further study of effective multivalent vaccines based on rGTPV. This study also provides a theoretical basis for overcoming the problem of low expression of exogenous genes.

Keywords

Goat pox virus Bidirectional promoter Vector Omp31 

Notes

Acknowledgments

This study was supported by the International Science and Technology Cooperation Project of China (2013DFR30970), the National Science and Technology Major Project (2013BAI05B05), the National Natural Science Foundation of China (U1303283, 31360610, 31602080), and the University Key Research Project of Henan Province (16A230013).

References

  1. Baldick CJ Jr, Keck JG, Moss B (1992) Mutational analysis of the core, spacer, and initiator regions of vaccinia virus intermediate-class promoters. J Virol 66:4710–4719PubMedPubMedCentralGoogle Scholar
  2. Berhe G et al (2003) Development of a dual recombinant vaccine to protect small ruminants against peste-des-petits-ruminants virus and capripoxvirus infections. J Virol 77:1571–1577CrossRefPubMedPubMedCentralGoogle Scholar
  3. Boyle DB (1992) Quantitative assessment of poxvirus promoters in fowlpox and vaccinia virus recombinants. Virus Genes 6:281–290CrossRefPubMedGoogle Scholar
  4. Caro-Hernández P et al (2007) Role of the Omp25/Omp31 family in outer membrane properties and virulence of Brucella ovis. Infect Immun 75:4050–4061CrossRefPubMedPubMedCentralGoogle Scholar
  5. Cochran MA, Puckett C, Moss B (1985) In vitro mutagenesis of the promoter region for a vaccinia virus gene: evidence for tandem early and late regulatory signals. J Virol 54:30–37PubMedPubMedCentralGoogle Scholar
  6. Cui M et al (2013) Impact of Hfq on global gene expression and intracellular survival in Brucella melitensis. PLoS ONE 8:e71933. doi: 10.1371/journal.pone.0071933 CrossRefPubMedPubMedCentralGoogle Scholar
  7. Davison AJ, Moss B (1989a) Structure of vaccinia virus early promoters. J Mol Biol 210:749–769CrossRefPubMedGoogle Scholar
  8. Davison AJ, Moss B (1989b) Structure of vaccinia virus late promoters. J Mol Biol 210:771–784CrossRefPubMedGoogle Scholar
  9. Diallo A et al (2002) Goat immune response to capripox vaccine expressing the hemagglutinin protein of peste des petits ruminants. Ann N Y Acad Sci 969:88–91CrossRefPubMedGoogle Scholar
  10. Estein SM, Cassataro J, Vizcaíno N, Zygmunt MS, Cloeckaert A, Bowden RA (2003) The recombinant Omp31 from Brucella melitensis alone or associated with rough lipopolysaccharide induces protection against Brucella ovis infection in BALB/c mice. Microbes Infect 5:85–93CrossRefPubMedGoogle Scholar
  11. Heald J, Berg J (2014) Immortalization of bovine dendritic cell clones for use in Brucella immunology research. In: Oral presentation at the undergraduate research day 2014, University of Wyoming, University of Wyoming Campus, 26 April 2014, p 37Google Scholar
  12. Hosamani M, Mondal B, Tembhurne PA, Bandyopadhyay SK, Singh RK, Rasool TJ (2004) Differentiation of sheep pox and goat poxviruses by sequence analysis and PCR-RFLP of P32 gene. Virus Genes 29:73–80CrossRefPubMedGoogle Scholar
  13. Kumar S, Boyle DB (1990a) Activity of a fowlpox virus late gene promoter in vaccinia and fowlpox virus recombinants. Arch Virol 112:139–148CrossRefPubMedGoogle Scholar
  14. Kumar S, Boyle DB (1990b) A poxvirus bidirectional promoter element with early/late and late functions. Virology 179:151–158CrossRefPubMedGoogle Scholar
  15. Mackett M, Smith GL, Moss B (1992) Vaccinia virus: a selectable eukaryotic cloning and expression vector. 1982. Biotechnology 24:495–499PubMedGoogle Scholar
  16. Perrin A et al (2007) Recombinant capripoxviruses expressing proteins of bluetongue virus: evaluation of immune responses and protection in small ruminants. Vaccine 25:6774–6783. doi: 10.1016/j.vaccine.2007.06.052 CrossRefPubMedGoogle Scholar
  17. Rao T, Bandyopadhyay S (2000) A comprehensive review of goat pox and sheep pox and their diagnosis. Anim Health Res Rev 1:127–136CrossRefPubMedGoogle Scholar
  18. Romero CH et al (1993) Single capripoxvirus recombinant vaccine for the protection of cattle against rinderpest and lumpy skin disease. Vaccine 11:737–742CrossRefPubMedGoogle Scholar
  19. Romero CH, Barrett T, Chamberlain RW, Kitching RP, Fleming M, Black DN (1994) Recombinant capripoxvirus expressing the hemagglutinin protein gene of rinderpest virus: protection of cattle against rinderpest and lumpy skin disease viruses. Virology 204:425–429. doi: 10.1006/viro.1994.1548 CrossRefPubMedGoogle Scholar
  20. Romero CH, Barrett T, Kitching RP, Bostock C, Black DN (1995) Protection of goats against peste des petits ruminants with recombinant capripoxviruses expressing the fusion and haemagglutinin protein genes of rinderpest virus. Vaccine 13:36–40CrossRefPubMedGoogle Scholar
  21. Tulman ER et al (2002) The genomes of sheeppox and goatpox viruses. J Virol 76:6054–6061CrossRefPubMedPubMedCentralGoogle Scholar
  22. Venkatesan G, Balamurugan V, Yogisharadhya R, Kumar A, Bhanuprakash V (2012) Differentiation of sheeppox and goatpox viruses by polymerase chain reaction-restriction fragment length polymorphism. Virol Sin 27:353–359. doi: 10.1007/s12250-012-3277-2 CrossRefPubMedGoogle Scholar
  23. Wade-Evans AM et al (1996) Expression of the major core structural protein (VP7) of bluetongue virus, by a recombinant capripox virus, provides partial protection of sheep against a virulent heterotypic bluetongue virus challenge. Virology 220:227–231. doi: 10.1006/viro.1996.0306 CrossRefPubMedGoogle Scholar
  24. Weir JP, Moss B (1987) Determination of the transcriptional regulatory region of a vaccinia virus late gene. J Virol 61:75–80PubMedPubMedCentralGoogle Scholar
  25. Yan X-M et al (2012) An outbreak of sheep pox associated with goat poxvirus in Gansu province of China. Vet Microbiol 156:425–428CrossRefPubMedGoogle Scholar
  26. Zantinge JL, Krell PJ, Derbyshire JB, Nagy E (1996) Partial transcriptional mapping of the fowlpox virus genome and analysis of the EcoRI L fragment. J Gen Virol 77(Pt 4):603–614CrossRefPubMedGoogle Scholar
  27. Zhao Z et al (2012) RNA interference targeting virion core protein ORF095 inhibits Goatpox virus replication in Vero cells. Virol J 9:48. doi: 10.1186/1743-422X-9-48 CrossRefPubMedPubMedCentralGoogle Scholar
  28. Zhou T et al (2012) Phylogenetic analysis of Chinese sheeppox and goatpox virus isolates. Virol J 9:25. doi: 10.1186/1743-422X-9-25 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Hui Zhang
    • 1
  • Zhihua Sun
    • 1
  • Na Zhang
    • 1
  • Zhiqiang Li
    • 2
  • Pengyan Wang
    • 1
  • Qiang Fu
    • 1
  • Yan Ren
    • 3
  • Xuehua Shao
    • 1
  • Yu Zhang
    • 1
  • Zhiru Guo
    • 1
  • Chuangfu Chen
    • 1
    Email author
  1. 1.College of Animal Science and TechnologyShihezi UniversityShihezi, Xinjiang ProvincePeople’s Republic of China
  2. 2.School of Life SciencesShangqiu Normal UniversityShangqiu, Henan ProvincePeople’s Republic of China
  3. 3.School of MedicineShihezi UniversityShihezi, Xinjiang ProvincePeople’s Republic of China

Personalised recommendations