Display options
Cite
Ericson BL, Carlson DJ, Carlson KA. Characterization of Nora Virus Structural Proteins via Western Blot Analysis. Scientifica (Cairo). 2016;2016:9067848doi: 10.1155/2016/9067848.
Ericson, B. L., Carlson, D. J., & Carlson, K. A. (2016). Characterization of Nora Virus Structural Proteins via Western Blot Analysis. Scientifica, 20169067848. https://doi.org/10.1155/2016/9067848
Ericson, Brad L, et al. "Characterization of Nora Virus Structural Proteins via Western Blot Analysis." Scientifica vol. 2016 (2016): 9067848. doi: https://doi.org/10.1155/2016/9067848
Ericson BL, Carlson DJ, Carlson KA. Characterization of Nora Virus Structural Proteins via Western Blot Analysis. Scientifica (Cairo). 2016;2016:9067848. doi: 10.1155/2016/9067848. Epub 2016 May 19. PMID: 27298753; PMCID: PMC4889860.
Copy Download .nbib Format: NLM
Share it on

Scientifica (Cairo). 2016;2016:9067848. doi: 10.1155/2016/9067848. Epub 2016 May 19.

Characterization of Nora Virus Structural Proteins via Western Blot Analysis.

Scientifica

Brad L Ericson, Darby J Carlson, Kimberly A Carlson

Affiliations

  1. Biology Department, University of Nebraska at Kearney, 2401 11th Avenue, Kearney, NE 68849, USA.

PMID: 27298753 PMCID: PMC4889860 DOI: 10.1155/2016/9067848

Abstract

Nora virus is a single stranded RNA picorna-like virus with four open reading frames (ORFs). The coding potentials of the ORFs are not fully characterized, but ORF3 and ORF4 are believed to encode the capsid proteins (VP3, VP4a, VP4b, and VP4c) comprising the virion. To determine the polypeptide composition of Nora virus virions, polypeptides from purified virus were compared to polypeptides detected in Nora virus infected Drosophila melanogaster. Nora virus was purified from infected flies and used to challenge mice for the production of antisera. ORF3, ORF4a, ORF4b, and ORF4c were individually cloned and expressed in E. coli; resultant recombinant proteins purified and were used to make monospecific antisera. Antisera were evaluated via Western blot against whole virus particles and Nora virus infected fly lysates. Viral purification yielded two particle types with densities of ~1.31 g/mL (empty particles) and ~1.33 g/mL (complete virions). Comparison of purified virus polypeptide composition to Nora virus infected D. melanogaster lysate showed the number of proteins in infected cell lysates is less than purified virus. Our results suggest the virion is composed of 6 polypeptides, VP3, VP4a, two forms of VP4b, and two forms of VP4c. This polypeptide composition is similar to other small RNA insect viruses.

References

  1. J Gen Virol. 1977 Feb;34(2):269-83 - PubMed
  2. J Virol. 1980 Jan;33(1):1-9 - PubMed
  3. J Gen Virol. 2006 Oct;87(Pt 10):3045-51 - PubMed
  4. Methods. 2001 Dec;25(4):402-8 - PubMed
  5. RNA Biol. 2013 Apr;10(4):481-9 - PubMed
  6. Virol J. 2008 Jul 18;5:82 - PubMed
  7. Nat Protoc. 2015 Jul;10(7):1084-97 - PubMed
  8. Dev Comp Immunol. 2014 Jan;42(1):67-84 - PubMed
  9. J Gen Virol. 2004 Jun;85(Pt 6):1497-507 - PubMed
  10. Virus Res. 2013 Aug;175(2):95-100 - PubMed
  11. J Virol. 1984 Jul;51(1):254-8 - PubMed
  12. Virus Res. 2011 Sep;160(1-2):51-8 - PubMed
  13. J Virol. 2004 Mar;78(6):3063-71 - PubMed

Publication Types

Grant support