Generation of a minigenome with non-coding sequences of infectious pancreatic necrosis virus

Abstract

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Infectious pancreatic necrosis virus (IPNV) is an important aquatic pathogen causing a highly devastating disease in salmonids. The virus has been isolated from over 50 species across the world. For combating the disease, vaccines have been developed by different recombinant DNA technologies. Production of live virus vaccines with defined attenuations requires reverse genetics system and minigenome synthesis to study the attenuation and virus production <i style="">in vitro</i> systems. Towards this objective, the two open reading frames of the IPNV (West Buxton strain) were genetically engineered to replace them with bacterial chloramphenicol acetyl transferase (CAT) reporter gene while retaining the non-coding regions (NCR). The minigenome of IPNV without the coding regions was generated using a modified pUC 19 plasmid and was checked for the nucleotide correctness by dideoxy chain termination method. Expression of the reporter gene was verified after transfection studies in susceptible cell line. The synthesised minigenome is useful in carrying out a number of studies in the reverse genetics of IPNV.

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