Construction of recombinant S-layer proteins (rSbsA) and their expression in bacterial ghosts - a delivery system for nontypeable Haemophilus influenzae antigen Omp26

Jennelle Kyd, K Jalava, W Lubitz

    Research output: Contribution to journalArticle

    28 Citations (Scopus)

    Abstract

    This study has investigated the feasibility of a combination of recombinant surface layer (S-layer) proteins and empty bacterial cell envelopes (ghosts) to deliver candidate antigens for a vaccine against nontypeable Haemophilus influenzae (NTHi) infections. The S-layer gene sbsA from Bacillus stearothermophilus PV72 was used for the construction of fusion proteins. Fusion of maltose binding protein (MBP) to the N-terminus of SbsA allowed expression of the S-layer in the periplasm of Escherichia coli. The outer membrane protein (Omp) 26 of NTHi was inserted into the N-terminal and C-terminal regions of SbsA. The presence of the fused antigen Omp26 was demonstrated by Western blot experiments using anti-Omp26 antisera. Electron microscopy showed that the recombinant SbsA maintained the ability to self-assemble into sheet-like and cylindrical structures. Recombinant E. coli cell envelopes (ghosts) were produced by the expression of SbsA/Omp26 fusion proteins prior to gene E-mediated lysis. Intraperitoneal immunization with these recombinant bacterial ghosts induced an Omp26-specific antibody response in BALB/c mice. These results demonstrate that the NTHi antigen, Omp26, was expressed in the S-layer self-assembly product and this construct was immunogenic for Omp26 when administered to mice in bacterial cell envelopes.
    Original languageEnglish
    Pages (from-to)185-192
    Number of pages8
    JournalPathogens and Disease
    Volume37
    Issue number2-3
    DOIs
    Publication statusPublished - 2003

    Fingerprint

    Haemophilus influenzae
    Recombinant Proteins
    Antigens
    Haemophilus Infections
    Maltose-Binding Proteins
    Escherichia coli
    Geobacillus stearothermophilus
    Periplasm
    Genes
    Antibody Formation
    Immune Sera
    Immunization
    Electron Microscopy
    Membrane Proteins
    Proteins
    Vaccines
    Western Blotting
    S-layer proteins

    Cite this

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    title = "Construction of recombinant S-layer proteins (rSbsA) and their expression in bacterial ghosts - a delivery system for nontypeable Haemophilus influenzae antigen Omp26",
    abstract = "This study has investigated the feasibility of a combination of recombinant surface layer (S-layer) proteins and empty bacterial cell envelopes (ghosts) to deliver candidate antigens for a vaccine against nontypeable Haemophilus influenzae (NTHi) infections. The S-layer gene sbsA from Bacillus stearothermophilus PV72 was used for the construction of fusion proteins. Fusion of maltose binding protein (MBP) to the N-terminus of SbsA allowed expression of the S-layer in the periplasm of Escherichia coli. The outer membrane protein (Omp) 26 of NTHi was inserted into the N-terminal and C-terminal regions of SbsA. The presence of the fused antigen Omp26 was demonstrated by Western blot experiments using anti-Omp26 antisera. Electron microscopy showed that the recombinant SbsA maintained the ability to self-assemble into sheet-like and cylindrical structures. Recombinant E. coli cell envelopes (ghosts) were produced by the expression of SbsA/Omp26 fusion proteins prior to gene E-mediated lysis. Intraperitoneal immunization with these recombinant bacterial ghosts induced an Omp26-specific antibody response in BALB/c mice. These results demonstrate that the NTHi antigen, Omp26, was expressed in the S-layer self-assembly product and this construct was immunogenic for Omp26 when administered to mice in bacterial cell envelopes.",
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    Construction of recombinant S-layer proteins (rSbsA) and their expression in bacterial ghosts - a delivery system for nontypeable Haemophilus influenzae antigen Omp26. / Kyd, Jennelle; Jalava, K; Lubitz, W.

    In: Pathogens and Disease, Vol. 37, No. 2-3, 2003, p. 185-192.

    Research output: Contribution to journalArticle

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