Skip to main content
SpringerLink
Log in

Functional analysis of RNAi suppressor P19 on improving baculovirus yield and transgene expression in Sf9 cells

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Objective

To investigate whether RNA interference suppressor P19 derived from tombusvirus can enhance baculovirus yield and transgene expression.

Results

A number of recombinant baculoviruses with P19 under the control of white spot syndrome virus immediate-early promoter, baculovirus early-to-late promoter, or P10 late promoter were constructed The budded virus titer and the expression levels of eGFP and luciferase were determined. P19 was clearly functional in Sf9 cells and could enhance baculovirus yield, eGFP and luciferase expression levels up to 6.8-, 1.8-, and 2.1-fold respectively, at 72 h post infection.

Conclusion

P19 enhanced baculovirus production and transgene expression, and thus has potential applications in baculovirus-based gene therapy and vaccine studies.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Berger I, Fitzgerald DJ, Richmond TJ (2004) Baculovirus expression system for heterologous multiprotein complexes. Nat Biotechnol 22:1583–1587

    Article  CAS  PubMed  Google Scholar 

  • Crawford AM, Miller LK (1988) Characterization of an early gene accelerating expression of late genes of the baculovirus Autographa californica nuclear polyhedrosis virus. J Virol 62:2773–2781

    PubMed Central  CAS  PubMed  Google Scholar 

  • Gao H, Wang Y, Li N, Peng WP, Sun Y, Tong GZ et al (2007) Efficient gene delivery into mammalian cells mediated by a recombinant baculovirus containing a whispovirus ie1 promoter, a novel shuttle promoter between insect cells and mammalian cells. J Biotechnol 131:138–143

    Article  CAS  PubMed  Google Scholar 

  • Jayachandran B, Hussain M, Asgari S (2012) RNA interference as a cellular defense mechanism against the DNA virus baculovirus. J Virol 86:13729–13734

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Kuzio J, Rohel DZ, Curry CJ, Krebs A, Carstens EB, Faulkner P (1984) Nucleotide sequence of the p10 polypeptide gene of Autographa californica nuclear polyhedrosis virus. Virology 139:414–418

    Article  CAS  PubMed  Google Scholar 

  • Lakatos L, Szittya G, Silhavy D, Burgyan J (2004) Molecular mechanism of RNA silencing suppression mediated by p19 protein of tombusviruses. EMBO J 23:876–884

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Li WX, Li H, Lu R, Li F, Dus M, Atkinson P et al (2004) Interferon antagonist proteins of influenza and vaccinia viruses are suppressors of RNA silencing. Proc Natl Acad Sci USA 101:1350–1355

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Mena JA, Kamen AA (2011) Insect cell technology is a versatile and robust vaccine manufacturing platform. Expert Rev Vaccines 10:1063–1081

    Article  CAS  PubMed  Google Scholar 

  • Oliveiraa VC, Bartassona L, Castrob ME, Corrêaa JR, Ribeiroa BM, Resendea RO (2011) A silencing suppressor protein (NSs) of a tospovirus enhances baculovirus replication in permissive and semipermissive insect cell lines. Virus Res 155:259–267

    Article  Google Scholar 

  • Qu F, Morris TJ (2002) Efficient infection of Nicotiana benthamiana by tomato bushy stunt virus is facilitated by the coat protein and maintained by p19 through suppression of gene silencing. Mol Plant Microbe Interact 15:193–202

    Article  CAS  PubMed  Google Scholar 

  • Rauschhuber C, Mueck-Haeusl M, Zhang WL, Nettelbeck DM, Ehrhardt A (2013) RNAi suppressor P19 can be broadly exploited for enhanced adenovirus replication and microRNA knock-down experiments. Sci Rep 3:1363

    Article  PubMed Central  PubMed  Google Scholar 

  • Siddiqui SA, Sarmiento C, Truve E, Lehto H, Lehto K (2008) Phenotypes and functional effects caused by various viral RNA silencing suppressors in transgenic Nicotiana benthamiana and N. tabacum. Mol Plant Microbe Interact 21:178–187

    Article  CAS  PubMed  Google Scholar 

  • van Rij RP, Saleh MC, Berry B, Foo C, Houk A, Antoniewski C, Andino R (2006) The RNA silencing endonuclease Argonaute 2 mediates specific antiviral immunity in Drosophila melanogaster. Genes Dev 20:2985–2995

    Article  PubMed Central  PubMed  Google Scholar 

  • Wang Y, Gao H, Chen C, Zhao HP, Li M, Sun Y et al (2008) Promoter activity of different promoters in recombinant Baculovirus-infected Sf9 cells. Chin J Biotechnol 24:598–603

    Google Scholar 

  • Wang W, Cheng T, Ma K, Xia D, Wang Y, Liu J et al (2013) Development of a novel baculovirus titration method using the enzyme-linked immunosorbent spot (ELISPOT) assay. J Virol Methods 188:114–120

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the Research Starting Foundation for Dr. Enqi Du, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (Grant No. SKLVBF201405) and National Natural Science Foundation of China (Grant No. 31201939), China.

Supporting information

Supplementary Table 1—Primers used.

Author information

Author notes

    Authors and Affiliations

    1. College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, People’s Republic of China

      Yangkun Liu, Yuanyuan Zhang, Dandan Liu, Enqi Du & Zengqi Yang

    2. China Institute of Veterinary Drug Control, Beijing, People’s Republic of China

      Lei Zhang

    Authors
    1. Yangkun Liu
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Lei Zhang
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Yuanyuan Zhang
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Dandan Liu
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Enqi Du
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Zengqi Yang
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding authors

    Correspondence to Enqi Du or Zengqi Yang.

    Additional information

    Yangkun Liu and Lei Zhang have contributed equally to this work.

    Electronic supplementary material

    Below is the link to the electronic supplementary material.

    Supplementary material 1 (DOCX 13 kb)

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Liu, Y., Zhang, L., Zhang, Y. et al. Functional analysis of RNAi suppressor P19 on improving baculovirus yield and transgene expression in Sf9 cells. Biotechnol Lett 37, 2159–2166 (2015). https://doi.org/10.1007/s10529-015-1910-5

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s10529-015-1910-5

    Keywords

    Search

    Navigation