Expression of cholera toxin epitopes inserted in flagellin by attenuated Salmonella strains

RESUMO

A novel antigen expression system has recently been developed by means of bacterial flagella, which are potent immunogens (Newton et al., Science 24470-72,1989). Here we show the insertion of two epitopes from the cholera toxin B-subunit in Salmonella flagelin, CTP-1 (residues 8-20) and CTP-3 (residues 50-64) (Jacob et al., Proc. Natl Acad. Sci. USA 81 7893-7896, 1984). First, we inserted CTP-1 for expression as a flagellar fusion; a hybrid flagellin expressing CTP-1 and CTP-3 in the hypervariable region of the flagellin gene was also constructed in an attempt to increase the cholera toxin neutralization potential, as well as to address some practical questions concerning the flagellar antigen expression system. The resulting constructs were non-motile although expression of chimeric flagellin was detected by immunoblotting and electron microscopy. One of the constructs (CTP-l + CTP-3) severely affected flagellin expression.