Structural and antigenic analysis of the yellow head virus nucleocapsid protein p20
| Autor: | Nusra Sittidilokratna, Peter J. Walker, Natthida Phetchampai, Vichai Boonsaeng |
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| Rok vydání: | 2005 |
| Předmět: |
Bacterial expression
Cancer Research Genes Viral Immunoblotting Molecular Sequence Data Shrimp virus Biology Roniviridae medicine.disease_cause Epitope Article Open Reading Frames Affinity chromatography Penaeidae Virology medicine Escherichia coli Amino Acid Sequence Isoelectric Point Amino Acids Cloning Molecular Peptide sequence Nucleoprotein chemistry.chemical_classification Nucleic acid sequence Yellow head virus Sequence Analysis DNA Nucleocapsid Proteins Molecular biology Recombinant Proteins Amino acid Molecular Weight Infectious Diseases chemistry Biochemistry Polyclonal antibodies biology.protein Epitopes B-Lymphocyte Epitope Mapping |
| Zdroj: | Virus Research |
| ISSN: | 0168-1702 |
| Popis: | Yellow head virus (YHV) is an invertebrate nidovirus that is highly pathogenic for marine shrimp. Nucleotide sequence analysis indicated that the YHV ORF2 gene encodes a basic protein (p I = 9.9) of 146 amino acids with a predicted molecular weight of 16,325.5 Da. The deduced amino acid sequence indicated a predominance of basic (15.1%), acidic (9.6%) and hydrophilic polar (34.3%) residues and a high proportion proline and glycine residues (16.4%). The ORF2 gene was cloned and expressed in Escherichia coli as a M r = 21 kDa His 6 -protein that reacted with YHV nucleoprotein (p20) monoclonal antibody. Segments representing the four linear quadrants of the nucleoprotein were also expressed in E. coli as GST-fusion proteins. Immunoblot analysis using YHV polyclonal rabbit antiserum indicated the presence of linear epitopes in all except the V 37 –Q 74 quadrant. Immunoblot analysis of the GST-fusion proteins and C-terminally truncated segments of the nucleoprotein allowed mapping of YHV monoclonal antibodies Y19, Y20 and YII4 to linear epitopes in the acidic domain between amino acids I 116 and E 137 . The full-length nucleoprotein was expressed at high level in E. coli and was easily purified in quantity from the soluble cell fraction by Ni + -NTA affinity chromatography. |
| Databáze: | OpenAIRE |
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