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Escherichia coli expression of NDV fusion protein gene and determination of its antigenic epitopes

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Abstract

Recurrent outbreaks of Newcastle disease have questioned the usage of existing vaccines that whether they are still adequate to protect clinical diseases and inhibit virus transmission in poultry. Advancement in molecular biology has led to the production of recombinant vaccines in recent years, which can be a more useful strategy to control infections of Newcastle disease virus (NDV). Studies indicate that the pathogenic nature of NDV is mediated by its membrane associated fusion (F) protein. Here we report the cloning of the full-length F gene-pET30a and its expression in Escherichia coli BL21 DE3 cells through isopropyl β-D-1-thiogalactopyranoside induction. Transferring the protein on nitrocellulose membrane in Western blotting confirmed its specificity with histidine-tagged antibody reaction at the proper size of 67 kDa. Protein purification with nickel charged sepharose column affinity chromatography resulted in a single band of 67 kDa purified His-tag F protein on SDS-PAGE. Analysis of its immunogenicity through bioinformatics tools revealed that more than 70% of its sequence is antigenically active comprising 24 linear immunogenic peptides predicted by the Linear epitope prediction tool and 9 immunogenic peptides predicted by ElliPro. This is a key achievement of the study, which may lead towards recombinant vaccine production in future. In conclusion, our findings suggest that rather than employing live viral vaccines, using a purified immunogenic recombinant F protein as a vaccine or cloning the same gene in a suitable plant vector for production of edible vaccine will provide better protection against the NDV into chicken.

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Abbreviations

F:

fusion protein

HN:

hemagglutinin neuraminidase

IPTG:

isopropyl β-D-1-thiogalactopyranoside

LB:

Luria Bertani

ND:

Newcastle disease

NDV:

Newcastle disease virus

PBS:

phosphate buffer saline

PI:

protrusion index

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Authors and Affiliations

  1. Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road, Lahore, 53700, Pakistan

    Naila Shahid, Sana Tahir, Abdul Q. Rao, Sameera Hassan, Anwar Khan, Ayesha Latif, Muhammad Au Khan, Bushra Tabassum, Ahmed A. Shahid, Ahmed U. Zafar & Tayyab Husnain

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  1. Naila Shahid
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  2. Sana Tahir
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  3. Abdul Q. Rao
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  4. Sameera Hassan
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  5. Anwar Khan
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  6. Ayesha Latif
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  7. Muhammad Au Khan
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  8. Bushra Tabassum
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  9. Ahmed A. Shahid
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  10. Ahmed U. Zafar
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  11. Tayyab Husnain
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Correspondence to Abdul Q. Rao.

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Electronic supplementary material. The online version of this article (DOI: https://doi.org/10.1515/biolog-2015-0191) contains supplementary material, which is available to authorized users.

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Shahid, N., Tahir, S., Rao, A.Q. et al. Escherichia coli expression of NDV fusion protein gene and determination of its antigenic epitopes. Biologia 70, 1553–1564 (2015). https://doi.org/10.1515/biolog-2015-0191

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  • DOI: https://doi.org/10.1515/biolog-2015-0191

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