PMID: 9636324Feb 1, 1996Paper

Development of new cloning vectors for the production of immunogenic outer membrane fusion proteins in Escherichia coli

Bio/technology
Pierre CornelisR Hamers

Abstract

The Pseudomonas aeruginosa lipoprotein gene (oprI) was modified by cloning an in-frame polylinker in both orientations at the end of oprI. The resulting plasmids pVUB1 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 OprI-based fusion proteins can be used to generate both humoral and cellular immune responses.

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Citations

Jun 1, 2002·Journal of Biotechnology·Patrik SamuelsonStefan Ståhl
Oct 14, 2003·FEMS Microbiology Letters·Su-Min KangJong-Won Oh
Aug 2, 2003·Research in Microbiology·Abdelaziz KholtiPierre Cornelis
Oct 12, 2000·Current Opinion in Biotechnology·P Cornelis
Jun 3, 2000·Nature Biotechnology·J S LeeC J Kim
Sep 15, 2001·Antimicrobial Agents and Chemotherapy·K E ConrathS Muyldermans
Jan 5, 2000·Applied and Environmental Microbiology·K KjaergaardP Klemm
Jul 11, 2002·Journal of Bacteriology·Kristian KjaergaardPer Klemm
Jul 25, 2014·Journal of Immunology Research·Afonso P Basto, Alexandre Leitão
Mar 19, 2014·Protein Expression and Purification·Afonso P BastoDulce M Santos
Dec 18, 2007·Immunity·Chiara MartinoliMaria Rescigno
Dec 4, 2014·Journal of Bioscience and Bioengineering·Shusuke OkamotoYuji Aso
Feb 12, 2015·FEMS Microbiology Reviews·Nienke Buddelmeijer
Jan 7, 2005·The Journal of Immunology : Official Journal of the American Association of Immunologists·Hilde RevetsPatrick De Baetselier
Sep 28, 1998·Journal of Bacteriology·C J LazdunskiH Bénédetti

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