Characterization of an immunoreactive region of the major capsid protein for infectious pancreatic necrosis virus Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/47429f465

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  • DNA fragments of the gene for VP2, the major capsid protein of Infectious Pancreatic Necrosis Virus (IPNV), were expressed in Escherichia coli and characterized immunologically with antisera made to different serotypes of IPNV. The entire VP2 gene was excised from the plasmid pUC19/A+SΔK and fragmented with restriction enzyme Sau3A1. The resulting fragments were inserted into the pATH vectors 1, 2, or 3 to form fusion proteins with the trpE protein which is under the control of the tryptophan promoter. Recombinants expressing the IPNV-VP2 antigens were detected by direct colony immunoblot with anti-IPNV sera and Western immunoblot analysis was used to further characterize the virus-specific proteins. A47 kDa reacting protein for the recombinant plasmid pB10/pB13 and a 52 kDa protein for pA43 were found. These proteins were serotypically distinct: pB10/pB13 was reactive with anti- IPNV/Sp, anti-IPNV/Buhl, and anti-IPNV/EVE sera; pA43 was only reactive with anti-IPNV/Sp sera, The nucleotide sequence of the pB10/pB13 insert was determined and mapped on the VP2 gene.
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