Cloning and expression of Treponema pallidum antigens in Escherichia coli.
A library of Treponema pallidum genomic DNA fragments produced by partial Sau3A digestion was established in Escherichia coli K12 using the plasmid vector pAT153. The library was screened using immune syphilitic rabbit serum and six recombinant phenotypes expressing eight treponemal polypeptides were detected. With two exceptions, all the recombinant gene products were the same size as polypeptides detected on Western immunoblots of T. pallidum. The genes encoding three novel gene products, with molecular masses in SDS-PAGE of 42, 17 and 15.5 kDa, which had not been cloned previously from T. pallidum were also identified. Monoclonal antibodies which reacted with four of the eight recombinant polypeptides were generated.
Bailey
M J
MJ
School of Biological Sciences, University of Birmingham, UK.
Thomas
C M
CM
Cockayne
A
A
Strugnell
R A
RA
Penn
C W
CW
D000911:Antibodies, Monoclonal / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics*; D015153:Blotting, Western; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics; D015183:Restriction Mapping; D014210:Treponema pallidum / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D004274:DNA, Recombinant