Rescue of recombinant Newcastle disease virus: current cloning strategies and RNA polymerase provision systems

2016 ◽  
Vol 162 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Aidin Molouki ◽  
Ben Peeters
2010 ◽  
Vol 5 (s1) ◽  
pp. e23-e24
Author(s):  
Bernardo Lozano-Dubernard ◽  
Ernesto Soto-Priante ◽  
David Sarfati-Mizrahi ◽  
Felipa Castro-Peralta ◽  
Ricardo Flores-Castro ◽  
...  

BioTechniques ◽  
2020 ◽  
Vol 68 (2) ◽  
pp. 96-100
Author(s):  
Pheik-Sheen Cheow ◽  
Tiong Kit Tan ◽  
Adelene Ai-Lian Song ◽  
Khatijah Yusoff ◽  
Suet Lin Chia

Reverse genetics has been used to generate recombinant Newcastle disease virus with enhanced immunogenic properties for vaccine development. The system, which involves co-transfecting the viral antigenomic plasmid with three helper plasmids into a T7 RNA polymerase-expressing cell to produce viral progenies, poses a great challenge. We have modified the standard transfection method to improve the transfection efficiency of the plasmids, resulting in a higher titer of virus progeny production. Two transfection reagents (i.e., lipofectamine and polyethylenimine) were used to compare the transfection efficiency of the four plasmids. The virus progenies produced were quantitated with flow cytometry analysis of the infectious virus unit. The modified transfection method increased the titer of virus progenies compared with that of the standard transfection method.


Virus Genes ◽  
2017 ◽  
Vol 53 (3) ◽  
pp. 410-417 ◽  
Author(s):  
Madhan Mohan Chellappa ◽  
Sohini Dey ◽  
Satish Gaikwad ◽  
Dinesh C. Pathak ◽  
Vikram N. Vakharia

2019 ◽  
Vol 14 (9) ◽  
pp. 617-628 ◽  
Author(s):  
Aidin Molouki ◽  
Abdou Nagy

Two decades have passed since the first reverse genetics system for the rescue of recombinant Newcastle disease virus was developed. Since then, the recombinant Newcastle disease virus vector has shown promising results as a safe and potent vector for development of many vaccines for both avian and human use. Herein, we review several technical topics that would be useful to further understanding of this technology. First, the effect of using helper plasmids encoding proteins belonging to strains other than the full-length cDNA and the possible incorporation of these expressed proteins into progeny virus will be discussed. Then, we will discuss the effect of removal of additional G residues from the T7 initiation sequence and finally, we will review different ways to improve rescue efficiency.


2014 ◽  
Vol 13 (2) ◽  
pp. 169-175 ◽  
Author(s):  
Yunzhou Wu ◽  
Shijun Yan ◽  
Zheng Lv ◽  
Lin Chen ◽  
Jingshu Geng ◽  
...  

2004 ◽  
Vol 78 (18) ◽  
pp. 10054-10063 ◽  
Author(s):  
Zhuhui Huang ◽  
Subbiah Elankumaran ◽  
Abdul S. Yunus ◽  
Siba K. Samal

ABSTRACT 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 to express the host-protective immunogen VP2 of a variant IBDV strain GLS-5. The gene encoding the VP2 protein of the IBDV was inserted into the most 3′-proximal locus of a full-length NDV cDNA for high-level expression. We successfully recovered the recombinant virus, rLaSota/VP2. The rLaSota/VP2 was genetically stable, at least up to 12 serial passages in chicken embryos, and was shown to express the VP2 protein. The VP2 protein was not incorporated into the virions of recombinant virus. Recombinant rLaSota/VP2 replicated to a titer similar to that of parental NDV strain LaSota in chicken embryos and cell cultures. To assess protective efficacy of the rLaSota/VP2, 2-day-old specific-pathogen-free chickens were vaccinated with the recombinant virus and challenged with a highly virulent NDV strain Texas GB or IBDV variant strain GLS-5 at 3 weeks postvaccination. Vaccination with rLaSota/VP2 generated antibody responses against both NDV and IBDV and provided 90% protection against NDV and IBDV. Booster immunization induced higher levels of antibody responses against both NDV and IBDV and conferred complete protection against both viruses. These results indicate that the recombinant NDV can be used as a vaccine vector for other avian pathogens.


Sign in / Sign up

Export Citation Format

Share Document