Rapid generation of rotavirus single-gene reassortants by means of eleven plasmid-only based reverse genetics
| Autor: | Riona Hatazawa, Koki Taniguchi, Takayuki Murata, Tetsushi Yoshikawa, Saori Fukuda, Yoshiki Kawamura, Satoshi Komoto |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Rotavirus
0301 basic medicine DNA Complementary Genotype Swine 030106 microbiology Reassortment Genome Viral Biology medicine.disease_cause Genome Rotavirus Infections Virus Cell Line 03 medical and health sciences Plasmid Cricetinae Virology medicine Animals Humans Gene Recombination Genetic Genetics Haplorhini Reverse Genetics Reverse genetics 030104 developmental biology RNA Viral Capsid Proteins Reassortant Viruses Plasmids |
| Zdroj: | Journal of General Virology. 101:806-815 |
| ISSN: | 1465-2099 0022-1317 |
| Popis: | Reassortment is an important mechanism in the evolution of group A rotaviruses (RVAs), yielding viruses with novel genetic and phenotypic traits. The classical methods for generating RVA reassortants with the desired genetic combinations are laborious and time-consuming because of the screening and selection processes required to isolate a desired reassortant. Taking advantage of a recently developed RVA reverse genetics system based on just 11 cloned cDNAs encoding the RVA genome (11 plasmid-only system), we prepared a panel of simian SA11-L2 virus-based single-gene reassortants, each containing 1 segment derived from human KU virus of the G1P[8] genotype. It was shown that there was no gene-specific restriction of the reassortment potential. In addition to these 11 single-gene reassortants, a triple-gene reassortant with KU-derived core-encoding VP1–3 gene segments with the SA11-L2 genetic background, which make up a virion composed of the KU-based core, and SA11-L2-based intermediate and outer layers, could also be prepared with the 11 plasmid-only system. Finally, for possible clinical application of this system, we generated a series of VP7 reassortants representing all the major human RVA G genotypes (G1–4, G9 and G12) efficiently. The preparation of each of these single-gene reassortants was achieved within just 2 weeks. Our results demonstrate that the 11 plasmid-only system allows the rapid and reliable generation of RVA single-gene reassortants, which will be useful for basic research and clinical applications. |
| Databáze: | OpenAIRE |
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