RESUMEN

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.

Asunto(s)

Proteínas de la Membrana Bacteriana Externa/genética , Proteínas Bacterianas/genética , Escherichia coli , Vectores Genéticos , Lipoproteínas/genética , Ingeniería de Proteínas , Vacunas Sintéticas , Virus de la Fiebre Porcina Africana/genética , Secuencia de Aminoácidos , Animales , Proteínas de la Membrana Bacteriana Externa/biosíntesis , Proteínas de la Membrana Bacteriana Externa/inmunología , Secuencia de Bases , Cápside/genética , Clonación Molecular , Genes Bacterianos , Lipoproteínas/biosíntesis , Datos de Secuencia Molecular , Proteínas Recombinantes de Fusión/biosíntesis