Anchisi S, etc.,al. Influenza A Virus Genetic Tools: From Clinical Sample to Molecular Clone. Methods Mol Biol. 2018;1836:33-58
Implementation of reverse genetics for influenza A virus, that is, the DNA-based generation of infectious viral particles in cell culture, opened new avenues to investigate the function of viral proteins and their interplay with host factors on a molecular level. This powerful technique allows the introduction, depletion, or manipulation of any given sequence in the viral genome, as long as it gives rise to replicating virus progeny. Reverse genetics can be used to generate targeted reassortant viruses by mixing segments of different viral strains, thus providing insight into phenotypes of potentially pandemic viruses arising from natural reassortment. It was further instrumental for the development of novel vaccine strategies, allowing rapid and targeted exchange of viral surface antigens on a well-replicating genetic backbone of cell culture-adapted or cold-adapted/attenuated viral strains. Establishment of reverse genetics and rescue of molecular clones of influenza A virus have been extensively described before. Here we give a detailed stand-alone protocol encompassing clinical sampling of influenza A virus specimens and subsequent plasmid-based genetics to rescue, manipulate, and confirm a fully infectious molecular clone. This protocol is based on the combined techniques and experience of a number of influenza laboratories, which are credited and referenced whenever appropriate.
See Also:
Latest articles in those days:
- Outbreak of Highly Pathogenic Avian Influenza A(H5N1) Virus in Seals, St. Lawrence Estuary, Quebec, Canada 2 days ago
- Molecular characterization of influenza virus circulating in Nepal in the year 2019 2 days ago
- Seasonal antigenic prediction of influenza A H3N2 using machine learning 2 days ago
- Probable extinction of influenza B/Yamagata and its public health implications: a systematic literature review and assessment of global surveillance databases 2 days ago
- Code to reproduce analysis in Nguyen et al, Emergence and interstate spread of highly pathogenic avian influenza A (H5N1) in dairy cattle. 2 days ago
[Go Top] [Close Window]