DEVELOPMENT OF NEW CLONING VECTORS FOR THE PRODUCTION OF IMMUNOGENIC OUTER-MEMBRANE FUSION PROTEINS IN ESCHERICHIA-COLI

The Pseudomonas aeruginosa lipoprotein gene (oprI) was modified by clo ning an in-frame polylinker in both orientations at the end of oprI. T he resulting plasmids pVUB1 and pVUB2 allow high lipoprotein productio n in E, coli after IPTG induction. The modified lipoproteins are prese nt in the outer membrane and surface-exposed. Outer membrane-bound fus ion proteins of different sizes were produced and used to generate ant ibodies without use of adjuvant. An 87 bp DNA fragment from the vp72 c apsid 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 ant igen-presenting cell line to a T-cell hybridoma while the correspondin g cross-linked S2b peptide was not. The results suggest that OprI-base d fusion proteins can be used to generate both humoral and cellular im mune responses.