Baculovirus expression of the glycoprotein gene of Lassa virus and characterization of the recombinant protein

A recombinant baculovirus was constructed that expresses the glycoprotein gene of Lassa virus (Josiah strain) under the transcriptional control of the polyhedrin promoter. The expressed protein (B-LSGPC) comigrated with the authentic viral glycoprotein as observed by sodium dodecyl sulfate polyacryl...

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Published inVirus research Vol. 25; no. 1-2; p. 79
Main Authors Hummel, K B, Martin, M L, Auperin, D D
Format Journal Article
LanguageEnglish
Published Netherlands 01.09.1992
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Abstract A recombinant baculovirus was constructed that expresses the glycoprotein gene of Lassa virus (Josiah strain) under the transcriptional control of the polyhedrin promoter. The expressed protein (B-LSGPC) comigrated with the authentic viral glycoprotein as observed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), was reactive with monoclonal antibodies (MAbs) in Western blots, and was glycosylated. Although the recombinant protein was not processed into the mature glycoproteins, G1 and G2, it demonstrated reactivity with all known epitopes as measured by indirect immunofluorescence (IFA), and it was immunogenic, eliciting antisera in rabbits that recognized whole virus in IFAs. Regarding future applications to diagnostic assays, the recombinant glycoprotein proved to be an effective substitute for Lassa virus-infected mammalian cells in IFAs and it was able to distinguish sera from several human cases of Lassa fever, against a panel of known negative sera of African origin, in an enzyme immunoassay (EIA).
AbstractList A recombinant baculovirus was constructed that expresses the glycoprotein gene of Lassa virus (Josiah strain) under the transcriptional control of the polyhedrin promoter. The expressed protein (B-LSGPC) comigrated with the authentic viral glycoprotein as observed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), was reactive with monoclonal antibodies (MAbs) in Western blots, and was glycosylated. Although the recombinant protein was not processed into the mature glycoproteins, G1 and G2, it demonstrated reactivity with all known epitopes as measured by indirect immunofluorescence (IFA), and it was immunogenic, eliciting antisera in rabbits that recognized whole virus in IFAs. Regarding future applications to diagnostic assays, the recombinant glycoprotein proved to be an effective substitute for Lassa virus-infected mammalian cells in IFAs and it was able to distinguish sera from several human cases of Lassa fever, against a panel of known negative sera of African origin, in an enzyme immunoassay (EIA).
Author Auperin, D D
Martin, M L
Hummel, K B
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Snippet A recombinant baculovirus was constructed that expresses the glycoprotein gene of Lassa virus (Josiah strain) under the transcriptional control of the...
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StartPage 79
SubjectTerms Animals
Antigens, Viral - genetics
Baculoviridae - genetics
Cell Line
Gene Expression
Genes, Viral
Glycoproteins - genetics
Glycoproteins - immunology
Humans
Immunoenzyme Techniques
Lassa Fever - diagnosis
Lassa virus - genetics
Lassa virus - immunology
Recombinant Proteins - genetics
Recombinant Proteins - immunology
Viral Proteins - genetics
Viral Proteins - immunology
Title Baculovirus expression of the glycoprotein gene of Lassa virus and characterization of the recombinant protein
URI https://www.ncbi.nlm.nih.gov/pubmed/1413995
Volume 25
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