Research Article Open Access

Cloning and Expression of the Nucleoprotein Gene (NP) of Newcastle Disease Virus (NDV) in Escherichia coli for Immunodiagnosis Application

Authors

CORRESPONDING AUTHOR
Ketherson Rodrigues Silva
Faculty of Agriculture and veterinay Sciences, Univ Estadual Paulista Jaboticabal Sao Paulo, Brazil
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Mariana Costa Mello Goncalves
Faculty of Agriculture and veterinay Sciences, Univ Estadual Paulista Jaboticabal Sao Paulo, Brazil
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Elisabete Schirato de Oliveira
Faculty of Agriculture and veterinay Sciences, Univ Estadual Paulista Jaboticabal Sao Paulo, Brazil
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Filipe Santos Fernando
Faculty of Agriculture and veterinay Sciences, Univ Estadual Paulista Jaboticabal Sao Paulo, Brazil
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Maria de Fatima Silva Montassier
Faculty of Agriculture and veterinay Sciences, Univ Estadual Paulista Jaboticabal Sao Paulo, Brazil
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Camila Cesario Fernandes
Faculty of Agriculture and veterinay Sciences, Univ Estadual Paulista Jaboticabal Sao Paulo, Brazil
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Maria de Lourdes Feres Tamanine
Faculty of Agriculture and veterinay Sciences, Univ Estadual Paulista Jaboticabal Sao Paulo, Brazil
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Mariana Monezi Borzi
Faculty of Agriculture and veterinay Sciences, Univ Estadual Paulista Jaboticabal Sao Paulo, Brazil
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Romeu Moreira dos Santos
Faculty of Agriculture and veterinay Sciences, Univ Estadual Paulista Jaboticabal Sao Paulo, Brazil
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Andre de Oliveira Mendonca
Faculty of Agriculture and veterinay Sciences, Univ Estadual Paulista Jaboticabal Sao Paulo, Brazil
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Dilmara Reischak
Faculty of Agriculture and veterinay Sciences, Univ Estadual Paulista Jaboticabal Sao Paulo, Brazil
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Antonio Carlos Paulillo
Faculty of Agriculture and veterinay Sciences, Univ Estadual Paulista Jaboticabal Sao Paulo, Brazil
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Helio Jose Montassier
Faculty of Agriculture and veterinay Sciences, Univ Estadual Paulista Jaboticabal Sao Paulo, Brazil
Published on: August 15, 2014
DOI 10.3923/ijps.2014.473.479
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Abstract

The nucleocapsid protein (NP) of newcastle disease virus (NDV) is an important antigen to develop a serologic assay on account of its highly conserved sequences and high immunogenicity. This study aimed to express the gene of the NP of NDV in a heterologous system (Escherichia coli), using the appropriate vector. The NDV-NP protein was expressed as a fusion recombinant protein containing SUMO peptide and poly-histidine tags. This recombinant nucleocapsid protein (rNP) was expressed in a soluble form which was easily purified and showed the ability to react with chicken anti-NDV polyclonal antibodies. An indirect ELISA method based on the adsorption of an antigen composed by NP (rNP-NDV-ELISA) was developed. By comparing this rNP-NDV-ELISA with haemagglutination-inhibition test (HI) high and significant correlation with the HI (r = 0.83) was found. In addition, high sensitivity (88.9%), specificity (95.5%), accuracy (90.4%) and agreement (0.85) were obtained. In conclusion the results indicated that the cloning and expression procedures used in this study provided a rNP that shared the major epitopes with the homologous viral protein and has the potential to be applied in ELISA for the immunodiagnosis of the Newcastle Disease.

Keywords:

Cloning of NDV-NP gene, expression, ELISA, immunodiagnosis Application, newcastle disease virus, recombinant nucleocapsid protien, SUMO

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How to Cite

Silva , K. R., Goncalves, M. C. M., Oliveira, E. S. de, Fernando, F. S., Montassier, M. de F. S., Fernandes, C. C., Tamanine, M. de L. F., Borzi, M. M., Santos, R. M. dos, Mendonca, A. de O., Reischak, D., Paulillo, A. C., & Montassier, H. J. (2014). Cloning and Expression of the Nucleoprotein Gene (NP) of Newcastle Disease Virus (NDV) in Escherichia coli for Immunodiagnosis Application. International Journal of Poultry Science, 13(8), 473–479. https://doi.org/10.3923/ijps.2014.473.479
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