An Efficient Method for Generating Poxvirus Recombinants in the Absence of Selection
Abstract
:1. Introduction
2. Results and Discussion
2.1. Generation of VV in FPV Permissive cells
2.2. Generation of Recombinant VV
2.3. The Use of Cells Non-Permissive for FPV to Rescue VV
2.4. Analysis of Resulting Viruses for Unintended Mutations and Pathogenicity
3. Experimental Section
3.1. Cell Culture
3.2. Virus Propagation
3.3. DNA Isolation from Virus
3.4. Generation of PCR Fragments
3.5. Verification of gfp/LacZ in TK Locus
3.6. Infection/Transfection
3.7. FPV Growth Curve
3.8. Animal Infection and Monitoring
3.9. RFLP Analysis of Vaccinia DNA
4. Conclusions
Acknowledgements
References and Notes
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Virus Isolate | Phenotype in Mice | CV-1:PK-15 Plaquing Efficiency Ratio |
---|---|---|
RwtVV 1 | <2 logs attenuated | 80 |
RwtVV 2 | <2 logs attenuated | 79074 |
RwtVV 3 | wild type | 21 |
RwtVV 4 | <2 logs attenuated | 275 |
RwtVV 5 | 1 log attenuated | 20 |
RwtVV 6 | wild type | 16 |
RwtVV 7 | 2 logs attenuated | 1084 |
RwtVV 8 | 1 log attenuated | 9074 |
RwtVV 9 | 1 log attenuated | 36 |
wtVV | wild type | 23 |
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Rice, A.D.; Gray, S.A.; Li, Y.; Damon, I.; Moyer, R.W. An Efficient Method for Generating Poxvirus Recombinants in the Absence of Selection. Viruses 2011, 3, 217-232. https://doi.org/10.3390/v3030217
Rice AD, Gray SA, Li Y, Damon I, Moyer RW. An Efficient Method for Generating Poxvirus Recombinants in the Absence of Selection. Viruses. 2011; 3(3):217-232. https://doi.org/10.3390/v3030217
Chicago/Turabian StyleRice, Amanda D., Stacey A. Gray, Yu Li, Inger Damon, and Richard W. Moyer. 2011. "An Efficient Method for Generating Poxvirus Recombinants in the Absence of Selection" Viruses 3, no. 3: 217-232. https://doi.org/10.3390/v3030217