Analysis of Haemophilus influenzae type b lipooligosaccharide-synthesis genes that assemble or expose a 2-keto-3-deoxyoctulosonic acid epitope

Y. Abu Kwaik, R. E. McLaughlin, M. A. Apicella, Stanley Spinola

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

We recently isolated a recombinant phage from a Haemophilus influenzae type b (Hib) library that assembles an oligosaccharide with an apparent molecular weight of 1400 (1.4K) on a 4.1K Escherichia coli lipopolysaccharide (LPS) structure, producing a 5.5K LPS species that contains a KDO (2-keto-deoxyoctulosonic acid) epitope. Subcloning and deletional analysis of the 14kb Haemophilus insert showed that three overlapping restriction fragments contained within a 7.2kb Pstl-BamHl fragment sequentially modified an E. coli 4.1K LPS structure, generating novel species of 4.5K, 5.1K and 5.5K. Only the 5.5K species contained the KDO epitope. We confirmed the relationship between the cloned genes and Hib lipooligosaccharide (LOS) biosynthesis by constructing a mutant that expressed an altered LOS. Thus, the Hib 7.2kb Pstl-BamHl restriction fragment contained a cluster of at least three genetic loci whose products acted sequentially in LOS biosynthesis.

Original languageEnglish (US)
Pages (from-to)2475-2480
Number of pages6
JournalMolecular Microbiology
Volume5
Issue number10
StatePublished - Oct 1991
Externally publishedYes

Fingerprint

Haemophilus influenzae type b
Keto Acids
Lipopolysaccharides
Epitopes
Escherichia coli
Genes
Haemophilus
Genetic Loci
Oligosaccharides
Bacteriophages
Libraries
Molecular Weight
lipid-linked oligosaccharides
2-keto-3-deoxyoctonate

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

Analysis of Haemophilus influenzae type b lipooligosaccharide-synthesis genes that assemble or expose a 2-keto-3-deoxyoctulosonic acid epitope. / Abu Kwaik, Y.; McLaughlin, R. E.; Apicella, M. A.; Spinola, Stanley.

In: Molecular Microbiology, Vol. 5, No. 10, 10.1991, p. 2475-2480.

Research output: Contribution to journalArticle

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