Molecular characterization and expression in Escherichia coli of the gene complex encoding the polysaccharide capsule of Neisseria meningitidis group B

Abstract

The gene complex encoding all determinants of the biosynthesis pathway of the capsule of group B meningococci (cps) has been cloned in Escherichia coli. A 24-kilobase large chromosomal fragment is necessary for capsule expression on the E. coli surface. By transposon and deletion mutagenesis, two separate steps in transport of the polysaccharide from the cytoplasm to the periplasm and further to the cell surface became evident. Mutants were also isolated that accumulate soluble poly(sialic acid) in the cytoplasm. The cloned cps complex conferred to E. coli strain GC6 sensitivity for E. coli K1-specific phages; phage sensitivity was enhanced in two distinct classes of cps mutants. Southern blot experiments revealed homology to some or all other Neisseria meningitidis capsular types and other Neisseria species, depending on the fragment of the cps complex used as probe.