Cloning and expression of viral haemorrhagic septicaemia nucleocapsid (N) gene of Rainbow trout (Onchorhynchus mykiss) in Escherichia coli
Septicemia viral disease (VHS) is one of the most important diseases of fish that has recently been developed in Iran and has caused severe damage to breeders. In order to identify the possible cause of the losses, farmed fish were sampled from epicenter where massive mortalities and economic losses had been occurred. Samples were collected from relevant tissues (i.e. liver, kidney, spleen, heart, and brain) of rainbow trout with clinical signs of VHS disease. The samples were stored in 70% ethanol, to perform RT-PCR. Then, recombinant plasmids were tested for protein expression in E. coli Rosetta strain. SDS-PAGE analysis indicated the production of a recombinant protein with an expected molecular weight of 61KDa. Thereafter, the recombinant protein was purified by amylose resin and its antigenicity was accessed by immunoblotting using a mouse polyclonal serum prepared against MBP protein. Positive reaction of the expressed protein with anti-MBP protein indicated that the expressed N protein possess antigenic epitopes and could be applied in future immunological studies.
Iranian Journal of Veterinary Clinical Sciences, Volume:12 Issue:2, 2019
97 - 108
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