Expression of a 32 kilodalton Theileria sergenti piroplasm surface protein by recombinant baculoviruses.
Previous studies detected a single amino acid substitution (Ala196 to Gly196) between cDNA clones encoding a 32 kDa antigen (p32) of Theileria sergenti (Chitose stock) obtained from a persistently infected calf. In this study, 2 different recombinant baculoviruses (pAc/p32-Ala196 and pAc/p32-Gly196) were constructed for the expression of p32. Molecular masses of the polypeptides produced in Spodoptera frugiperda cells infected with the recombinant baculoviruses were the same as that of authentic p32. pAc/p32-Ala196 produced additional polypeptides, with molecular masses higher than 32 kDa, which resulted from differential N-glycosylation as revealed by endo N-glycosidase treatment. The results indicate that a single amino acid substitution may lead to a conformational change in p32 which affected post-translational modification of recombinant products.
Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
T
T
Matsuba
C
C
Sugimoto
M
M
Hattori
Y
Y
Sako
K
K
Fujisaki
M
M
Onuma
D000409:Alanine; D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D005998:Glycine; D015151:Immunoblotting; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017354:Point Mutation; D015800:Protozoan Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D018411:Spodoptera; D016794:Theileria / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D018076:DNA, Complementary; D008565:Membrane Proteins; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C085175:piroplasm surface protein p32, Theileria sergenti; D000409:Alanine; D005998:Glycine