Characterization of Nora Virus Structural Proteins via Western Blot Analysis
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 com...
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Wiley
2016-01-01
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| Series: | Scientifica |
| Online Access: | http://dx.doi.org/10.1155/2016/9067848 |
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| author | Brad L. Ericson Darby J. Carlson Kimberly A. Carlson |
| author_facet | Brad L. Ericson Darby J. Carlson Kimberly A. Carlson |
| author_sort | Brad L. Ericson |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-221f14117c1f4e058ec447a62684f626 |
| institution | Kabale University |
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| language | English |
| publishDate | 2016-01-01 |
| publisher | Wiley |
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| spelling | doaj-art-221f14117c1f4e058ec447a62684f6262025-02-03T01:26:11ZengWileyScientifica2090-908X2016-01-01201610.1155/2016/90678489067848Characterization of Nora Virus Structural Proteins via Western Blot AnalysisBrad L. Ericson0Darby J. Carlson1Kimberly A. Carlson2Biology Department, University of Nebraska at Kearney, 2401 11th Avenue, Kearney, NE 68849, USABiology Department, University of Nebraska at Kearney, 2401 11th Avenue, Kearney, NE 68849, USABiology Department, University of Nebraska at Kearney, 2401 11th Avenue, Kearney, NE 68849, USANora 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.http://dx.doi.org/10.1155/2016/9067848 |
| spellingShingle | Brad L. Ericson Darby J. Carlson Kimberly A. Carlson Characterization of Nora Virus Structural Proteins via Western Blot Analysis Scientifica |
| title | Characterization of Nora Virus Structural Proteins via Western Blot Analysis |
| title_full | Characterization of Nora Virus Structural Proteins via Western Blot Analysis |
| title_fullStr | Characterization of Nora Virus Structural Proteins via Western Blot Analysis |
| title_full_unstemmed | Characterization of Nora Virus Structural Proteins via Western Blot Analysis |
| title_short | Characterization of Nora Virus Structural Proteins via Western Blot Analysis |
| title_sort | characterization of nora virus structural proteins via western blot analysis |
| url | http://dx.doi.org/10.1155/2016/9067848 |
| work_keys_str_mv | AT bradlericson characterizationofnoravirusstructuralproteinsviawesternblotanalysis AT darbyjcarlson characterizationofnoravirusstructuralproteinsviawesternblotanalysis AT kimberlyacarlson characterizationofnoravirusstructuralproteinsviawesternblotanalysis |