Abstract
Plasmids pQE-60 and pQE-30 containing 6 x His-tag sequence were used for expression of fragments of HCV structural and non-structural proteins in Escherichia coli (E. coli). The following fragments were used: core (1-98 aa), NS3 (202-482 aa), and tetramer of hypervariable region 1 (HVR1) of E2 protein. The constructed plasmids directed high levels of expression of HCV proteins in E. coli JM109. After purification by the metal-affinity chromatography on nickel-nitrilotriacetic acid (Ni-NTA) agarose, the His-tagged HCV proteins were used for immunization of BALB/c mice. All three proteins were able to induce high levels of specific antibodies and, in the case of the NS3 and HVR1 tetramer, also to mount vigorous cell-proliferating responses. High immunogenicity of the tested fragments of HCV proteins shows them as good candidates for inclusion into the future HCV vaccine preparations.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antigen-Antibody Reactions
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Cell Proliferation
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Chromatography, Affinity / methods
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Cloning, Molecular
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Dose-Response Relationship, Immunologic
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Female
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Hepatitis C Antibodies / immunology
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Hepatitis C Antigens / genetics
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Hepatitis C Antigens / immunology
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Hepatitis C Antigens / isolation & purification*
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Immunization
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Mice
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Mice, Inbred BALB C
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Nickel / chemistry
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Nitrilotriacetic Acid / chemistry
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Peptide Fragments / genetics
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Peptide Fragments / immunology
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Peptide Fragments / isolation & purification*
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Peptides / genetics
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Peptides / immunology
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Peptides / isolation & purification*
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Recombinant Proteins / genetics
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Recombinant Proteins / immunology
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Recombinant Proteins / isolation & purification
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Sensitivity and Specificity
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Spleen / cytology
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Spleen / immunology
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Viral Core Proteins / genetics
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Viral Core Proteins / immunology
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Viral Core Proteins / isolation & purification*
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Viral Nonstructural Proteins / genetics
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Viral Nonstructural Proteins / immunology
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Viral Nonstructural Proteins / isolation & purification*
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Viral Proteins / genetics
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Viral Proteins / immunology
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Viral Proteins / isolation & purification*
Substances
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HVR1 protein, Hepatitis C virus
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Hepatitis C Antibodies
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Hepatitis C Antigens
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NS3 protein, hepatitis C virus
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Peptide Fragments
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Peptides
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Recombinant Proteins
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Viral Core Proteins
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Viral Nonstructural Proteins
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Viral Proteins
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core protein (1-98), hepatitis C virus
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core protein p22, Hepatitis C virus
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Nickel
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Nitrilotriacetic Acid