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| Content Provider | Springer Nature : BioMed Central |
|---|---|
| Author | Xu, Qingyuan Ge, Jinying Li, Maolin Sun, Encheng Zhou, Yawei Guo, Yunze Wu, Donglai Bu, Zhigao |
| Abstract | Background Bluetongue virus (BTV), an emerging insect vector mediated pathogen affecting both wild ruminants and livestock, has a genome consisting of 10 linear double-stranded RNA genome segments. BTV has a severe economic impact on agriculture in many parts of the world. Current reverse genetics (RG) strategy to rescue BTV mainly rely on in vitro synthesis of RNA transcripts from cloned complimentary DNA (cDNA) corresponding to viral genome segments with the aid of helper plasmids. RNA synthesis is a laborious job which is further complicated with a need for expensive reagents and a meticulous operational procedure. Additionally, the target genes must be cloned into a specific vector to prepare templates for RNA transcription. Result In this study, we have developed a PCR based BTV RG system with easy two-step transfection. Viable viruses were recovered following a first transfection with the seven helper plasmids and a second transfection with the 10 PCR products on the BSR cells. Further, recovered viruses were characterized with indirect immunofluorescence assays (IFA) and gene sequencing. And the proliferation properties of these viruses were also compared with wild type BTV. Interestingly, we have identified that viruses containing the segment 2 of the genome from reassortant BTV, grew slightly slower than the others. Conclusion In this study, a convenient PCR based RG platform for BTV is established, and this strategy could be an effective alternative to the original available BTV rescue methods. Furthermore, this RG strategy is likely applicable for other Orbiviruses. |
| Related Links | https://virologyj.biomedcentral.com/counter/pdf/10.1186/s12985-019-1261-2.pdf |
| Ending Page | 9 |
| Page Count | 9 |
| Starting Page | 1 |
| File Format | HTM / HTML |
| DOI | 10.1186/s12985-019-1261-2 |
| Journal | Virology Journal |
| Issue Number | 1 |
| Volume Number | 16 |
| Language | English |
| Publisher | BioMed Central |
| Publisher Date | 2019-12-05 |
| Access Restriction | Open |
| Subject Keyword | Virology Bluetongue virus Reverse genetics T7 RNA polymerase Genome modification Reassortment |
| Content Type | Text |
| Resource Type | Article |
| Subject | Virology Infectious Diseases |
| Journal Impact Factor | 4/2023 |
| 5-Year Journal Impact Factor | 3.8/2023 |
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