Expression of the S1 sub-unit of pertussis toxin using a recombinant baculovirus.
The full-length genome of the S1 sub-unit containing the native signal sequence was inserted downstream of the polyhedrin promoter of Autographa californica nuclear polyhedrosis virus. A large amount of 28 Kd protein, which was larger than the authentic S1 sub-unit (26 Kd), was detected in insect cells infected with the recombinant virus by Coomassie blue staining after resolving by SDS-PAGE. Although the level of expression of the recombinant S1 sub-unit was high, immunoblotting and N-terminal sequencing analyses of the expressed product revealed that the signal sequence of the S1 was poorly processed in the insect cells. In vitro assembly of the recombinant S1 sub-unit with native B oligomer to form holotoxin was low, based on an estimation of both cell cytotoxicity against Chinese hamster ovary (CHO) cells and binding activity to haptoglobin and anti-B oligomer antibody.
Matsuura
Y
Y
Department of Veterinary Science, National Institute of Health, Tokyo, Japan.
Sato
H
H
Sato
Y
Y
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002478:Cells, Cultured; D015870:Gene Expression; D005822:Genetic Vectors; D037342:Pertussis Toxin; D011487:Protein Conformation; D015202:Protein Engineering; D011995:Recombination, Genetic; D010566:Virulence Factors, Bordetella / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin