Construction of Recombinant Bacmid DNA Encoding Newcastle Disease Virus (NDV) Fusion Protein Gene

Authors

  • Majid Moghbeli Department of Microbiology, Islamic Azad University, Damaghan Branch, Damaghan, Iran
  • Mohammad reza Shafaati Department of Microbiology, Islamic Azad University, Damaghan Branch, Damaghan, Iran
  • Roholah Dorostkar Applied Virology Research Center, Department of Virology, Baqiyatalah University of Medical Sciences, Tehran, Iran
Abstract:

Background and Aims: Newcastle disease virus (NDV) is one of the major pathogen in poultry. Vaccination is intended to control the disease as an effective solution nevertheless this virus is a growing threat to the poultry industry. F gene open reading frame (ORF) from NDV is 1650 bp, encoding a protein of 553 amino acids that can induce protective immunity alone. The F glycoprotein on the surface of NDV is important for virus infectivity and pathogenicity. Towards protection goal, the full-length of F gene was isolated using specific primers and cloned into the baculovirus derived bacmid shuttle vector to produce recombinant F-protein in insect cells. Materials and Methods: F gene ORF from avirulent strain La Sota of NDV was amplified by RT-PCR to produce F cDNA. The amplicon was cloned firstly into the T/A cloning vector and then subcloned into the pFastBac Dual donor plasmid through NcoI/KpnI sites. After the verification of cloning process by PCR and enzymatic digestion analysis, the accuracy of F gene ORF in the T/A cloning vector was confirmed by sequencing. Finally, F-containing recombinant bacmid was subsequently generated and confirmed by PCR using F specific primers and M13 universal primers. Results: Results showed that a recombinant baculovirus containing a correct and in framework sequence of Newcastle F gene under the control of p10 promoter was constructed. Conclusion: The above mentioned F-containing recombinant baculovirus, in addition to its independent application, can be used with other individual recombinant baculoviruses expressing NH and NP genes to produce Newcastle VLPs in insect cell line.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

construction of recombinant bacmid dna encoding newcastle disease virus (ndv) fusion protein gene

background and aims: newcastle disease virus (ndv) is one of the major pathogen in poultry. vaccination is intended to control the disease as an effective solution nevertheless this virus is a growing threat to the poultry industry. f gene open reading frame (orf) from ndv is 1650 bp, encoding a protein of 553 amino acids that can induce protective immunity alone. the f glycoprotein on the surf...

full text

construction of recombinant bacmid dna encoding influenza virus a (h1n1) hemagglutinin gene

objective: influenza virus a (h1n1) is an important subtype of the influenza respiratory viruses, which has important worldwide implications. hemagglutinin (ha), an important viral antigen, is responsible for binding to human cell receptors leading to an onset of the disease process. considering the critical role of viral attachment, this study focuses on the extraction and cloning of ha and it...

full text

A recombinant Newcastle disease virus (NDV) expressing VP2 protein of infectious bursal disease virus (IBDV) protects against NDV and IBDV.

Infectious bursal disease virus (IBDV) causes a highly immunosuppressive disease in chickens. Currently available, live IBDV vaccines can lead to generation of variant viruses. We have developed an alternative vaccine that will not create variant IBDV. By using the reverse genetics approach, we devised a recombinant Newcastle disease virus (NDV) vector from a commonly used vaccine strain LaSota...

full text

Construction of a recombinant bacmid DNA containing influenza A virus hemagglutinin gene using a site-specific transposition mechanism

Introduction: In recent years, influenza viruses have caused moderate to severe infections all around the world while so far there is no influenza vaccine that can protect people with only one dose of injection. In this regard, producing a universal vaccine based on virus-like-particles (VLP) could be an ideal approach.  Methods: In this study, the full-length ORF of influenza hemagglutini...

full text

Determination of immune potentials of recombinant fusion and recombinant haemagglutinin-neuraminidase antigens of Newcastle Disease virus (NDV)

Poultry sector is one of the major components of Indian animal husbandry and provides continuous and consistent income around the year to the poultry farmers. Further, animal products like meat and egg also bring foreign revenue into the country. The growing demand for acceptance in the global market for poultry products warrants ISSN: 2319-7706 Volume 4 Number 1 (2015) pp. 575-588 http://www.i...

full text

Perspective on Possible Recombination Event in Fusion Protein Gene of Newcastle Disease Viruses Isolated in Iran

Background and Aims: Newcastle disease (ND), caused by the virulent Newcastle disease virus (NDV), is one of the most important viral diseases in birds. In recent years recombination occurring throughout the NDVs genome isolated in China and Indonesia has been reported. This study was focused to investigate the recombination events in the F gene of the Iranian NDVs to generate useful data that ...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 7  issue None

pages  15- 20

publication date 2013-07

By following a journal you will be notified via email when a new issue of this journal is published.

Keywords

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023