Tn916-generated, lipooligosaccharide mutants of Neisseria meningitidis and Neisseria gonorrhoeae.

Infection and Immunity
David S StephensM A Apicella

Abstract

A library of Tn916-generated, tetracycline-resistant (Tc) mutants of the group B Neisseri meningitidis strain NMB was screened by using monoclonal antibodies (MAbs) that recognize structural differences in neisserial lipooligosaccharide (LOS). The LOS of parental strain NMB had a relative molecular mass of 4.5 kDa, reacted with MAbs 3F11 and 6B4 but not with MAb 4C4 or 6E4, and contained a lacto-N-neotetrose unit. Two phenotypically stable mutants, SS3 and R6, altered in LOS, were identified by colony immunoblots, electrophoresis, and Western immunoblots. The LOS of mutant SS3 was 3.4 kDa and reacted with MAbs 4C4 and 6E4 but not MAb 3E11 or 6B4. The LOS of mutant R6 was 3.1 to 3.2 kDa and reacted with MAb 6E4 but not MAb 3F11, 6B4, or 4C4. Thus, the LOSs of the R6 and SS3 mutants were predicted to contain different truncations of the core oligosaccharide. The LOS phenotype of each mutant was linked to Tc(r), as determined by transformation of the parent strain with DNA from the mutant. Southern hybridizations and single-specific-primer PCR revealed in each mutant a single truncated tn916 insertion which had lost genes required for mobilization. Tn916 mutagenesis was used to identify two distinct genetic sites in the meningococ...Continue Reading

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Citations

May 18, 2005·The Journal of Experimental Medicine·Rachel M ExleyChristoph M Tang
Jan 15, 1999·Critical Reviews in Microbiology·C M Kahler, D S Stephens
Jun 11, 1999·Molecular Microbiology·G C ShihD S Stephens
Jul 20, 2005·The Journal of Antimicrobial Chemotherapy·Holli LancasterPeter Mullany
Sep 15, 2000·Journal of Bacteriology·V PelicicX Nassif
Mar 6, 1998·Infection and Immunity·M M EstabrookM A Apicella
Jan 5, 2002·Journal of Microbiological Methods·John A BogdanM S Blake
Jul 8, 2000·Microbes and Infection·Y L Tzeng, D S Stephens

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