The Membrane-Anchoring Region of the AcMNPV P74 Protein Is Expendable or Interchangeable with Homologs from Other Species
Abstract
:1. Introduction
2. Materials and Methods
2.1. In Vitro Cell Culture and Insect Rearing
2.2. Bioinformatics Studies
2.3. P74 Gene Knockout in AcMNPV-bacmid
2.4. AcMNPV-bacmidΔp74 Complementation
2.5. OB Generation for the Recombinant Baculoviruses
2.6. Bioassays with Recombinant AcMNPVs
3. Results
3.1. P74 Phylogeny and Protein Domains
3.1.1. Phylogeny
3.1.2. Protein Domains
3.2. AcMNPV Knockout in p74 Gene and Complementation
3.2.1. CRISPR/Cas9 Application
3.2.2. AcMNPV OBs with P74 Variants
3.3. Infectivity of AcMNPV Variants
3.3.1. Infectivity of AcMNPV Variants in S. exigua
3.3.2. Infectivity of AcMNPV Variants in R. nu
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Primer | Sequence (5′ to 3′) |
---|---|
FwP74seq | GGTTTTAACAGCCGTCGATTTA |
RvP74seq | TAAGATGCATTTGTTGTCGAGC |
pFwp74Ac | GAAGATCTATGGCGGTTTTAACAG |
pRevp74Ac | CCCAAGCTTAAAATAACAAATCAATTG |
pRevBamAc | TAACCGAACGGATCCCATAGCGC |
pFwBamAc | GCGCTATGGGATCCGTTCGGTTA |
pFwP74Se | ATCAGATCTGCTATGCTCACTTTTGTAGAC |
pRvP74Se | ATCAAGCTTATTCCGAATAGAGATTGTCGTAC |
pFwP74Ha | ATCAGATCTTTCATGTCGAATATCATTTATATAC |
pRvP74Ha | ATCAAGCTTATGTGTATAAATTGTGGTACC |
FwMe53 | GCTTTGAAATGCACAACG |
P74 | AcMNPV | SeMNPV | HearSNPV | |
---|---|---|---|---|
AcMNPV | 100 | 55 | 52 | |
Complete | SeMNPV | 82 | 100 | 55 |
HearSNPV | 82 | 82 | 100 | |
AcMNPV | 100 | 59 | 57 | |
Nt | SeMNPV | 85 | 100 | 59 |
HearSNPV | 83 | 84 | 100 | |
AcMNPV | 100 | 46 | 42 | |
Ct | SeMNPV | 78 | 100 | 46 |
HearSNPV | 77 | 77 | 100 |
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Nugnes, M.V.; Targovnik, A.M.; Mengual-Martí, A.; Miranda, M.V.; Cerrudo, C.S.; Herrero, S.; Belaich, M.N. The Membrane-Anchoring Region of the AcMNPV P74 Protein Is Expendable or Interchangeable with Homologs from Other Species. Viruses 2021, 13, 2416. https://doi.org/10.3390/v13122416
Nugnes MV, Targovnik AM, Mengual-Martí A, Miranda MV, Cerrudo CS, Herrero S, Belaich MN. The Membrane-Anchoring Region of the AcMNPV P74 Protein Is Expendable or Interchangeable with Homologs from Other Species. Viruses. 2021; 13(12):2416. https://doi.org/10.3390/v13122416
Chicago/Turabian StyleNugnes, María Victoria, Alexandra Marisa Targovnik, Adrià Mengual-Martí, María Victoria Miranda, Carolina Susana Cerrudo, Salvador Herrero, and Mariano Nicolás Belaich. 2021. "The Membrane-Anchoring Region of the AcMNPV P74 Protein Is Expendable or Interchangeable with Homologs from Other Species" Viruses 13, no. 12: 2416. https://doi.org/10.3390/v13122416
APA StyleNugnes, M. V., Targovnik, A. M., Mengual-Martí, A., Miranda, M. V., Cerrudo, C. S., Herrero, S., & Belaich, M. N. (2021). The Membrane-Anchoring Region of the AcMNPV P74 Protein Is Expendable or Interchangeable with Homologs from Other Species. Viruses, 13(12), 2416. https://doi.org/10.3390/v13122416