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Development of New Cloning Vectors for the Production of Immunogenic Outer Membrane Fusion Proteins in Escherichia coli

Bio/Technology volume 14, pages 203–208 (1996)Cite this article

Abstract

The Pseudomonas aeruginosa lipoprotein gene (oprl) was modified by cloning an in-frame polylinker in both orientations at the end of oprl. The resulting plasmids pVUBl and pVUB2 allow high lipoprotein production in E. coli after IPTG induction. The modified lipoproteins are present in the outer membrane and surface-exposed. Outer membrane-bound fusion proteins of different sizes were produced and used to generate antibodies without use of adjuvant. An 87 bp DNA fragment from the vp72 capsid protein gene of African Swine Fever virus (ASFV) and the entire Leishmania major glycoprotein gp63 gene were expressed in this system. Finally, a fusion lipoprotein containing a 16 amino acid epitope from the pre-S2b region of Hepatitis B virus (HBV) was presented by an antigen-presenting cell line to a T-cell hybridoma while the corresponding cross-linked S2b peptide was not. The results suggest that Oprl-based fusion proteins can be used to generate both humoral and cellular immune responses.

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Authors and Affiliations

  1. Laboratorium Algemene Biologie, Vrije Universiteit Brussel, Vlaams Instituut Biotechnologie, Paardenstraat 65, B-1640, St. Genesius Rode, Belgium

    Pierre Cornells, Javier Cote Sierra, Antonio Lim Jr., Achut Malur, Sumalee Tungpradabkul & Hamida Tazka

  2. Laboratorio de Doenças Infecciosas, Faculdade de Medicina Veterinaria, Rua Gomes Freire, 1199, Lisboa, Portugal

    Alexandre Leitão, Carlos V. Martins & Raymond Hamers

  3. CVZ Instituto Investigação Cientifíca Tropical, Lisboa, Portugal

    Alexandre Leitão

  4. Laboratorium Cellulaire Immunologie, Vrije Universiteit Brussel, Vlaams Instituut Biotechnologie, Paardenstraat 65, B-1640, St. Genesius Rode, Belgium

    Christi di Perna, Lea Brys & Patrick De Baetselier

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  1. Pierre Cornells
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Correspondence to Pierre Cornells.

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Cornells, P., Sierra, J., Lim, A. et al. Development of New Cloning Vectors for the Production of Immunogenic Outer Membrane Fusion Proteins in Escherichia coli. Nat Biotechnol 14, 203–208 (1996). https://doi.org/10.1038/nbt0296-203

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