The biologic role of a majority of the Neisseria meningitidis 2100 predicted coding regions is still to be assigned or experimentally confirmed. Determining the phenotypic effect of gene disruption being a fundamental approach to understanding gene function, we used high-density signature-tagged transposon mutagenesis, followed by a large-scale sequencing of the transposon insertion sites, to construct a genome-wide collection of mutants. The sequencing results for the first half of the 4548 mutants composing the library suggested that we have mutations in 80%-90% of N. meningitidis nonessential genes. This was confirmed by a whole-genome identification of the genes required for resistance to complement-mediated lysis, a key to meningococcal virulence. We show that all the genes we identified, including four previously uncharacterized, were important for the synthesis of the polysialic acid capsule or the lipooligosaccharide (LOS), suggesting that these are likely to be the only meningococcal attributes necessary for serum resistance. Our work provides a valuable and lasting resource that may lead to a global map of gene function in N. meningitidis.
Large-scale analysis of the meningococcus genome by gene disruption: resistance to complement-mediated lysis.
M. Geoffroy,Stéphanie Floquet,Arnaud Métais,X. Nassif,V. Pelicic
Published 2003 in Genome Research
ABSTRACT
PUBLICATION RECORD
- Publication year
2003
- Venue
Genome Research
- Publication date
2003-03-01
- Fields of study
Biology, Medicine
- Identifiers
- External record
- Source metadata
Semantic Scholar, PubMed
CITATION MAP
EXTRACTION MAP
CLAIMS
CONCEPTS
- complement-mediated lysis
Destruction of bacterial cells by the host complement system, used here as a phenotype linked to virulence.
Aliases: complement lysis
- genes required for resistance to complement-mediated lysis
The subset of genes recovered from the screen that are associated with survival during complement attack.
Aliases: complement-resistance genes
- genome-wide mutant library
The pooled collection of transposon insertion mutants generated for systematic analysis of gene function in the organism.
Aliases: mutant library, mutant collection
- high-density signature-tagged transposon mutagenesis
A transposon-based mutagenesis approach used here to generate many tagged insertion mutants across the bacterial genome.
Aliases: signature-tagged transposon mutagenesis, HSTM
- large-scale sequencing of transposon insertion sites
A sequencing-based mapping approach used to identify where transposons inserted in the mutant collection.
Aliases: insertion site sequencing, transposon insertion site sequencing
- lipooligosaccharide (los)
A bacterial outer-membrane glycolipid whose biosynthesis affects meningococcal surface composition.
Aliases: LOS, lipooligosaccharide
- neisseria meningitidis
The meningococcal bacterial species whose genome and mutant collection are analyzed in the abstract.
Aliases: meningococcus, N. meningitidis
- nonessential genes
Genes that can be disrupted without preventing viability under the conditions tested in this mutagenesis screen.
Aliases: nonessential gene set
- polysialic acid capsule
A sialic-acid-containing bacterial capsule structure involved in meningococcal surface properties.
Aliases: capsule polysialic acid, polysialic capsule
- serum resistance
The ability of meningococci to survive exposure to complement-containing serum.
Aliases: complement resistance
REFERENCES
Showing 1-40 of 40 references · Page 1 of 1