Construction of Recombinant Rabies Virus Vectors Expressing H or F Protein of Peste des Petits Ruminants Virus
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Viruses, Cells, Serum, and Plasmids
2.2. Target Gene Synthesis
2.3. Amplification and Purification of the Target Gene
2.4. Construction and Verification of Recombinant Full-Length Infectious Clones
2.5. Rescue of Recombinant Virus
2.6. Direct Immunofluorescence and Indirect Immunofluorescence
2.7. Western Blot
2.8. Transmission Electron Microscopy (TEM)
2.9. Growth Kinetics of Recombinant RABV
2.10. Genetic Stability of Antigenic Genes in Recombinant RABV
3. Results
3.1. Construction and Identification of pD-SRV9-PM-PPRV-H and pD-SRV9-PM-PPRV-F
3.2. Direct Immunofluorescence Identification of the Successful Rescue of the Recombinant Viruses
3.3. Identification of Recombinant Virus Rescue via Indirect Immunofluorescence
3.4. Verification of the Correct Expression of Recombinant Virus H and F via Western Blot
3.5. TEM Revealed That the Recombinant Virus Particles Were Bullet-Shaped
3.6. Growth Kinetics of Recombinant Viruses
3.7. Stable Inheritance of Exogenous Gene Expression in Recombinant Viruses
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Primer | Sequence (5′–3′) | Enzye |
---|---|---|
PM/PPRV/H/F | TTTCGTACGGCCACCATGTCCGCACAAAGGGAAAGGATCA | PmeI |
PM/PPRV/H/R | ACAGTTTAAACTCAAACTGGATTGCATGTTACCTCT | BsiWI |
PM/PPRV/F/F | TTTCGTACGGCCACCATGACACGGGTCGCAATCTTGACAT | PmeI |
PM/PPRV/F/R | ACAGTTTAAACCTACAGTGATCTCACATACGACTTT | BsiwI |
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Wang, H.; Bi, J.; Feng, N.; Zhao, Y.; Wang, T.; Li, Y.; Yan, F.; Yang, S.; Xia, X. Construction of Recombinant Rabies Virus Vectors Expressing H or F Protein of Peste des Petits Ruminants Virus. Vet. Sci. 2022, 9, 555. https://doi.org/10.3390/vetsci9100555
Wang H, Bi J, Feng N, Zhao Y, Wang T, Li Y, Yan F, Yang S, Xia X. Construction of Recombinant Rabies Virus Vectors Expressing H or F Protein of Peste des Petits Ruminants Virus. Veterinary Sciences. 2022; 9(10):555. https://doi.org/10.3390/vetsci9100555
Chicago/Turabian StyleWang, Haojie, Jinhao Bi, Na Feng, Yongkun Zhao, Tiecheng Wang, Yuetao Li, Feihu Yan, Songtao Yang, and Xianzhu Xia. 2022. "Construction of Recombinant Rabies Virus Vectors Expressing H or F Protein of Peste des Petits Ruminants Virus" Veterinary Sciences 9, no. 10: 555. https://doi.org/10.3390/vetsci9100555