Construction and Rescue of a DNA-Launched DENV2 Infectious Clone
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Construction of Infectious Clone Plasmid
2.3. Preparation of Virus Stocks
2.4. Virus Infections
2.5. Immunoblots
2.6. Immunofluorescence Microscopy
2.7. Splicing Assay
2.8. Plaque Assay
2.9. Statistics
3. Results and Discussion
3.1. Design and Construction of a DNA-Launched DENV2 16681 Infectious Clone
3.2. Rescue of DENV2 16681
3.3. Characterization of Rescued DENV2 16681
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Name | Sequence (5′ to 3′) | Template |
---|---|---|
Vector_F | GTTGAATCAACAGGTTCTGGCCGGCATGGTCCCAGCCTC | pcDNA6.2-ZIKV MR766-HDVr |
Vector_R | CGTAGACTAACAACTCGGTTCACTAAACGAGCTCTGC | |
DENV 5′UTR | GCAGAGCTCGTTTAGTGAACCGAGTTGTTAGTCTACGTGGACCGACAAAGACAGATTCTTTGAGGGAGCTAAGCTCAACGTAGTTCTAACAGTTTTTTAATTAGAGAGCAGATCTCTGATGAATAACCAACGGAAAAAGGCG | gBlock |
PCR-A_F | GATCTCTGATGAATAACCAACGGAAAAAGGCG | pcDNA3.1 DENV 16681 CprME |
PCR-A_R | CACAACGCAACCACTATCGGCCTGCACCATGACTCCCAAATAC | |
NS1-Intron | GTATTTGGGAGTCATGGTGCAGGCCGATAGTGGTTGCGTTGTGAGCTGGAAAAACAAAGAACTGAAATGTGGCAGTGGGATTTTCATCACAGACAACGTGCACACATGGACAGAACAATACAAGTTCCAACCAGAATCCCCTTCAAAACTAGCTTCAGCTATCCAGAAAGTAAGTATCAAGGTTACAAGACAGGTTTAAGGAGACCAATAGAAACTGGGCTTGTCGAGACAGAGAAGACTCTTGCGTTTCTGATAGGCACCTATTGGTCTTACTGACATCCACTTTGCCTTTCTCTCCACAGGCCCATGAAGAGGGCATTTGTGGAATCCGCTCAGTAACAAGACTGGAGAATCTGATGTGGAAACAAATAACACCAGAATTGAATCACATTCTATCAGAAAATGAGGTGAAGTTAACTATTATGACAGGAGACATCAAAGGAATCATGCAGGCAGGAAAACGATCTCTGCGGCCTCAGCCC | gBlock |
PCR-B_F | GATCTCTGCGGCCTCAGCCCACTGAGCTGAAG | pcDNA3.1 DENVrepGFP/Zeo |
PCR-B_R | GGGACCATGCCGGCCAGAACCTGTTGATTCAACAGCACC |
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Holliday, M.; Corliss, L.; Lennemann, N.J. Construction and Rescue of a DNA-Launched DENV2 Infectious Clone. Viruses 2023, 15, 275. https://doi.org/10.3390/v15020275
Holliday M, Corliss L, Lennemann NJ. Construction and Rescue of a DNA-Launched DENV2 Infectious Clone. Viruses. 2023; 15(2):275. https://doi.org/10.3390/v15020275
Chicago/Turabian StyleHolliday, Madeline, Lochlain Corliss, and Nicholas J. Lennemann. 2023. "Construction and Rescue of a DNA-Launched DENV2 Infectious Clone" Viruses 15, no. 2: 275. https://doi.org/10.3390/v15020275