Scheme for generation of <i>N. meningitidis</i> LbpB mutant strains.

<p>PCR products of the indicated regions were amplified from plasmids isolated from yeast and used to transform <i>N. meningitidis</i> strain MC58 or its derivatives. The indicated regions were assembled in the yeast shuttle vector pYES2 (invitrogen) using the yeast homologous recombination method (Shao et al, 2009). In the first step, the <i>lbpB</i> gene is replaced by a chloramphenicol resistance gene. The resulting strain (N364) was transformed with PCR products that introduced wild-type (strain N368) or mutant <i>lbpB</i> genes (−LG or –LG, -SM; strains N365 and N366) followed by a gentamicin resistance cassette.