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Genetic mutant organisms pervade all areas of Biology. Early on, herpesviruses HV were found to be amenable to genetic analysis using homologous recombination techniques in eukaryotic cells. More recently, HV genomes cloned onto a bacterial artificial chromosome BAC have become available. HV BACs can be easily modified in E.coli and reintroduced in eukaryotic cells to produce infectious viruses. Mutants derived from HV BACs have been used both to understand the functions of all types of genetic elements present on the virus genome, but also to generate mutants with potentially medically relevant properties such as preventative vaccines. Here we retrace the development of the BAC technology applied to the Epstein-Barr virus EBV and review the strategies available for the construction of mutants. We expand on the appropriate controls required for proper use of the EBV BACs, and on the technical hurdles researchers face in working with these recombinants. We then discuss how further technological developments might successfully overcome these difficulties. Finally, we catalog the EBV BAC mutants that are currently available and illustrate their contributions to the field using a few representative examples.

Electronic supplementary materialThe online version of this article doi:10.1186-2042-4280-1-6 contains supplementary material, which is available to authorized users.

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Autor: Regina Feederle - Emmalene J Bartlett - Henri-Jacques Delecluse

Fuente: https://link.springer.com/

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