Escherichia coli expression of NDV fusion protein gene and determination of its antigenic epitopes
-
Naila Shahid
,
Sameera Hassan
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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Articles in the same Issue
- Escherichia coli expression of NDV fusion protein gene and determination of its antigenic epitopes
- Screening for fibrinolytic filamentous fungi and enzymatic properties of the most potent producer, Aspergillus brasiliensis AUMC 9735
- Biochemical, anatomical and molecular level changes in cucumber (Cucumis sativus) seedlings exposed to copper oxide nanoparticles
- Expression of bacterial glutamine synthetase gene in Arabidopsis thaliana increases the plant biomass and level of nitrogen utilization
- Comparison between natural and impacted Alpine lakes six years after hydropower exploitation has ceased
- Formation of radial symmetric needle-like rosette leaves in Arabidopsis
- Contribution to the knowledge of lohmanniid oribatid mites (Acari: Oribatida: Lohmanniidae) of Cuba
- Aquatic insects of a lowland rainforest in Papua New Guinea: assemblage structure in relation to habitat type
- Short-horned grasshopper subfamilies feed at different rates on big bluestem and switchgrass cultivars
- Nests of the harvest mouse (Micromys minutus) as habitat for invertebrates
- Comparison of the anatomical arrangement of the branches arising from the descending aorta in rabbit (Oryctolagus cuniculus f. domestica) and European hare (Lepus europaeus)
- Molecular dynamics simulation and docking studies on novel mutants (T11V, T12P and D364S) of the nucleotide-binding domain of human heat shock 70 kDa protein
- Paraoxonase 1 activity and polymorphisms in multiple sclerosis patients
