High level expression of the B. microplus Bm86 antigen in the yeast Pichia pastoris forming highly immunogenic particles for cattle.
Recently, a gene coding for the Bm86 tick gut glycoprotein was cloned, expressed in Escherichia coli and shown to induce an immunological response in cattle to damage ticks engorging on these animals (Rand et al., 1989). We report here the increased expression of the Bm86 antigen from the cattle tick Boophilus microplus in the methylotrophic yeast Pichia pastoris. The recombinant protein was obtained with a purity higher than 95% by a procedure with a high yield. The conducted biochemical studies demonstrated the antigen to be glycosylated and found to form particles of around 17 to 45 nm in diameter with enhanced immunogenic properties. Ticks engorging on vaccinated cattle were significantly damaged as a result of the immune response against the recombinant antigen. This system permits the obtainment in a high yield of the tick Bm86 antigen, in a glycosylated and particulated form.
Rodríguez
M
M
Mammalian Cell Genetics Division, Centro de Ingenieria Genética y Biotecnología, Havana, Cuba.
Rubiera
R
R
Penichet
M
M
Montesinos
R
R
Cremata
J
J
Falcón
V
V
Sánchez
G
G
Bringas
R
R
Cordovés
C
C
Valdés
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D011817:Rabbits; D011994:Recombinant Proteins; D013987:Ticks / Q000276:immunology*; D014611:Vaccination; D014612:Vaccines; D014614:Vaccines, Synthetic / Q000276:immunology*
C085065:BM86 protein, Boophilus; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D014612:Vaccines; D014614:Vaccines, Synthetic