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Genetic and antigenic characterization of recombinant nucleocapsid proteins derived from canine coronavirus and canine respiratory coronavirus in China
- Source :
- Science China. Life Sciences
- Publication Year :
- 2016
- Publisher :
- Springer Science and Business Media LLC, 2016.
-
Abstract
- To characterize the antigenicity of nucleocapsid proteins (NP) derived from canine coronavirus (CCoV) and canine respiratory coronavirus (CRCoV) in China, the N genes of CCoV (CCoV-BJ70) and CRCoV (CRCoV-BJ202) were cloned from swabs obtained from diseased pet dogs in Beijing and then sequenced. The recombinant NPs (rNPs) were expressed in Escherichia coli and purified by nickel-affinity column and size exclusion chromatography. Sequencing data indicated that the N genes of CCoV-BJ70 and CRCoV-BJ202 belonging to two distinctly different groups were relatively conserved within each subgroup. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) results showed that rNPs of CCoV and CRCoV were expressed efficiently and isolated with a final purity of over 95%. Western blot analysis revealed the rNP from CRCoV could cross-react with mice antisera against human coronavirus (HCoV-229E, NL63, OC43, HKU1), while rNP of CCoV had cross-reactivity with only anti-sera against viruses belonging to the same group (HCoV-229E and NL63). In summary, CCoV and CRCoV rNPs were successfully expressed in E. coli and showed antigenic cross-reactivity with antisera raised against human coronaviruses. These findings indicate that further serologic studies on coronavirus infections at the animal-human interface are needed.
- Subjects :
- 0301 basic medicine
China
Antigenicity
Genes, Viral
cross-reactivity
Blotting, Western
030106 microbiology
medicine.disease_cause
General Biochemistry, Genetics and Molecular Biology
law.invention
03 medical and health sciences
Dogs
Coronavirus, Canine
Western blot
Environmental Science(all)
law
expression
medicine
Animals
Amino Acid Sequence
Cloning, Molecular
canine respiratory coronavirus
Antigens, Viral
Escherichia coli
Gene
Polyacrylamide gel electrophoresis
Phylogeny
General Environmental Science
Antiserum
Sequence Homology, Amino Acid
Agricultural and Biological Sciences(all)
biology
medicine.diagnostic_test
Biochemistry, Genetics and Molecular Biology(all)
virus diseases
Canine coronavirus
Nucleocapsid Proteins
biology.organism_classification
Virology
canine coronavirus
030104 developmental biology
Recombinant DNA
Electrophoresis, Polyacrylamide Gel
General Agricultural and Biological Sciences
nucleocapsid protein
Research Paper
Subjects
Details
- ISSN :
- 18691889 and 16747305
- Volume :
- 59
- Database :
- OpenAIRE
- Journal :
- Science China Life Sciences
- Accession number :
- edsair.doi.dedup.....96402e10fe34b2dc4e267cb8b91e2b21