1. Development of new cloning vectors for the production of immunogenic outer membrane fusion proteins in Escherichia coli
- Author
-
Sierra Jc, Alexandre Leitão, Tazka H, Hamers R, di Perna C, Lea Brys, Tungpradabkul S, Pierre Cornelis, De Baetseller P, Carlos Martins, Antonio Jr. Lim, and Achut Malur
- Subjects
Lipoproteins ,Recombinant Fusion Proteins ,Genetic Vectors ,Molecular Sequence Data ,Biomedical Engineering ,Cloning vector ,Bioengineering ,Biology ,Molecular cloning ,Protein Engineering ,Applied Microbiology and Biotechnology ,Plasmid ,Capsid ,Bacterial Proteins ,Multiple cloning site ,Escherichia coli ,Animals ,Amino Acid Sequence ,Cloning, Molecular ,Vaccines, Synthetic ,Base Sequence ,Lipid bilayer fusion ,Fusion protein ,Molecular biology ,African Swine Fever Virus ,Membrane protein ,Genes, Bacterial ,Molecular Medicine ,Bacterial outer membrane ,Biotechnology ,Bacterial Outer Membrane Proteins - 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.
- Published
- 1996