Production and secretion of pertussis toxin subunits in Bacillus subtilis.
Pertussis toxin (PT) is a major component of today's acellular whooping cough vaccines. The use of acellular vaccines is predicted to increase sharply in the near future. There is therefore a need to produce PT in a way that makes its purification as easy as possible. Our approach was to express all five PT subunits individually in Bacillus subtilis. We have used vectors containing the promoter and signal sequences of the alpha-amylase gene of Bacillus amyloliquefaciens followed by an insert encoding the appropriate PT-subunit. All PT-subunits were secreted and found in the culture supernatant. The level of expression varied considerably: S1 and S5 were produced in large quantities whereas much smaller amounts of S2, S3 and S4 were found. The subunits were also present in the membrane fraction of the respective strains.
Saris
P
P
Department of Bacteriology, National Public Health Institute, Helsinki, Finland.
Taira
S
S
Airaksinen
U
U
Palva
A
A
Sarvas
M
M
Palva
I
I
Runeberg-Nyman
K
K
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D037342:Pertussis Toxin; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D010566:Virulence Factors, Bordetella / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism
D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin