Biochemical characterization of the recombinant Boophilus microplus Bm86 antigen expressed by transformed Pichia pastoris cells.
In the present paper we report the biochemical characteristics of the recombinant tick (Boophilus microplus) gut antigen Bm86 that previously has been cloned, expressed and recovered at high levels in the methylotrophic yeast Pichia pastoris. The results demonstrate that rBm86 had a modification at position 92 (Thr replaced by Ile) and aggregated, forming particles ranging between 17 and 40 nm. The rBm86 was N-glycosylated, having at least two non-glycosylated sequons (Asn-329 and Asn-363) and a ratio of only 0.4/65 (free Cys/total Cys)/mol of protein.
Montesino
R
R
Division of Industrial Biotechnology, Centro de Ingeniería Genética y Biotechnología, La Habana, Cuba.
Cremata
J
J
Rodríguez
M
M
Besada
V
V
Falcón
V
V
de la Fuente
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000941:Antigens / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D002461:Cell Line, Transformed; D006031:Glycosylation; D037102:Lectins; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010316:Particle Size; D010843:Pichia; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013987:Ticks / Q000276:immunology*
D000941:Antigens; D037102:Lectins; D011994:Recombinant Proteins