Molecular Cloning and Expression of the Fusion (F) Gene from Newcastle Disease Virus in Escherichia coli: A Platform for Further Studies

Document Type : Original Articles

Authors

1 Department of Biology, Faculty of Sciences, Shahid Chamran University of Ahvaz, Ahvaz, Iran.

2 Department of Pathobiology, School of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran.

3 Department of Animal Sciences, Agricultural and Natural Resources University of khuzestan, Iran

10.32592/ARI.2025.80.3.715

Abstract

Newcastle disease (ND), is a highly contagious viral disease, affecting most of the avian species. The fusion protein in the ND virus serves as the target for immune response. The goal of this study was to develop the DNA vaccine using a fusion gene from the Newcastle virus. A new candidate DNA vaccine against Newcastle disease virus (NDV) has been developed. This innovative vaccine uses a fusion gene that encodes immunogenic proteins derived from NDV. The hypothesis behind this approach is that the fusion gene induces a strong immune response against the virus, potentially leading to long-term immunity in vaccinated individuals. Fusion gene RNA was extracted from the Newcastle virus, amplified by the reverse transcription-polymerase chain reaction (RT-PCR). After that, it was sub-cloned in the pTG-19T vector and then in expression vector pET43.1a E. coli BL21. Gene expression was induced by IPTG. The fusion protein was subjected to Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Sequencing and PCR findings confirmed the cloning of the fusion gene into the vector. Digestion results showed the target gene had been inserted in the pET43.1a plasmid, successfully. SDS-PAGE revealed a protein band of about 54.7 kDa. Analysis of the constructs in E.coli cells revealed the successful expression of gene inserts in vitro. Our results show that the fusion protein produced by pET43.1a in E. coli can be used as a DNA vaccine. However, a weak band of expressed protein was found and the fusion protein produced by pET43.1a in E. coli was not so efficient. This survey encourages researchers to do more studies for testing the produced protein as a vaccine in vivo and in vitro.

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Archives of Razi Institute
Volume 80, Issue 3
May and June 2025
Pages 715-720

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