Evaluation of a Subunit Vaccine to Infectious Hematopoietic Necrosis Virus, July 31, 1989 to September 30, 1990, Annual Report.

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The IHNV glycoprotein has been identified as the virion protein which elicits neutralizing antibody in rabbits and induces protective immunity in fish to homologous and heterologous strains of IHNV (Engelking and Leong, 1989). These findings suggested that genetic engineering might be used to develop an economically feasible IHNV vaccine for fish. Thus, a clone of the IHNV glycoprotein gene was made and expression of a portion of this gene in bacteria resulted in a prototype IHNV subunit vaccine. Only 350 bases of IHNV sequence was expressed in this initial vaccine construction because there were 16 cysteine residues in the glycoprotein ... continued below

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77 pages

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Leong, JoAnn Ching September 1, 1990.

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Description

The IHNV glycoprotein has been identified as the virion protein which elicits neutralizing antibody in rabbits and induces protective immunity in fish to homologous and heterologous strains of IHNV (Engelking and Leong, 1989). These findings suggested that genetic engineering might be used to develop an economically feasible IHNV vaccine for fish. Thus, a clone of the IHNV glycoprotein gene was made and expression of a portion of this gene in bacteria resulted in a prototype IHNV subunit vaccine. Only 350 bases of IHNV sequence was expressed in this initial vaccine construction because there were 16 cysteine residues in the glycoprotein gene. Previous work with the rabies glycoprotein had shown that when the entire gene was expressed in bacteria, a denatured protein was produced, presumably because appropriate folding mechanisms for disulfide bond formation in protein were absent in E. coli. The IHNV vaccine clone contained a region of the gene which encoded only one cysteine residue. Despite the efficacy of the vaccine in laboratory trials, it seemed useful to determine whether other regions of the IHNV glycoprotein gene would be expressed in an antigenically recognizable form in bacteria and thereby, provide increased protection in fish. The recombinant plasmids pXL2, pXL3, and pXL7 were constructed so that all regions of the glycoprotein gene were expressed in bacteria as trpE-G fusion proteins. All of these recombinant plasmids produced fusion proteins that were also analyzed in Western immunoblots with anti-IHNV sera and specific monoclonal antibodies. These results were compared with the proteins produced by p52G and p618G, the plasmids identified in the original vaccine construction. The results of this comparison are shown.

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77 pages

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  • Other Information: PBD: 1 Sep 1990

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  • Report No.: DOE/BP-16479-4
  • Grant Number: 1984BP16479
  • DOI: 10.2172/758124 | External Link
  • Office of Scientific & Technical Information Report Number: 758124
  • Archival Resource Key: ark:/67531/metadc707404

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  • September 1, 1990

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  • Sept. 12, 2015, 6:31 a.m.

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Leong, JoAnn Ching. Evaluation of a Subunit Vaccine to Infectious Hematopoietic Necrosis Virus, July 31, 1989 to September 30, 1990, Annual Report., report, September 1, 1990; United States. (digital.library.unt.edu/ark:/67531/metadc707404/: accessed November 13, 2018), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.