Graduate Thesis Or Dissertation

 

Epitope mapping and characterization of the glycoprotein of infectious hematopoietic necrosis virus Public Deposited

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  • A characterization of the antigenic determinants (epitopes) of theglycoprotein (G) of infectious hematopoietic necrosis virus (IHNV) wasmade with different regions of the G gene expressed in Escherichia coli. AcDNA copy of the G gene was divided into four fragments after Taq Idigestion and these fragments were subcloned into the pATH vectorswhich put expression of each G gene fragment under the control of the trpEpromoter. The resulting plasmids encoded trpE-G fusion proteinscontaining different regions of the viral glycoprotein. The three plasmids
  • A characterization of the antigenic determinants (epitopes) of theglycoprotein (G) of infectious hematopoietic necrosis virus (IHNV) wasmade with different regions of the G gene expressed in Escherichia coli. AcDNA copy of the G gene was divided into four fragments after Taq Idigestion and these fragments were subcloned into the pATH vectorswhich put expression of each G gene fragment under the control of the trpEpromoter. The resulting plasmids encoded trpE-G fusion proteinscontaining different regions of the viral glycoprotein. The three plasmids,pXL2, pXL3, and pXL7, were found to encode fusion proteins that weredetected with anti-IHNV sera in Western immunoblots. A comparision ofreactivities of the fusion proteins encoded by these plasmids was madewith a number of anti-G specific monoclonal antibodies (Mabs) by Westernimmunoblot and radioimmuoassay. The non-neutralizing monoclonalantibody, 136J, was found to react with the trpE-G fusion proteinencoded by pXL3 and the fusion proteins encoded by plasmids, p52G andp618G, which were identified in previous studies (Gilmore et al., 1988). Another non-neutralizing Mab, 2F, was able to bind to the pXL3 fusionprotein and the neutralizing Mab, RB B5, recognized the pXL7 fusionprotein. Competitive radioimmune studies with a synthetic peptidederived from the amino acid sequence encoded by pXI_3 was found toinhibit the binding of a neutralizing Mab, 127B, to purified IHN virus.
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