Open AccessThe identification a novel gene required for lipopolysaccharide biosynthesis by Haemophilus influenzae RM7004, using transposon Tn916 mutagenesis
Author(s) -
High Nicola J.,
Deadman Mary E.,
Hood Derek W.,
Moxon E. Richard
Publication year - 1996
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1996.tb08596.x
Subject(s) - transposon mutagenesis , biology , transposable element , gene , mutagenesis , microbiology and biotechnology , escherichia coli , open reading frame , mutant , genetics , peptide sequence
Mutagenesis with the transposon Tn916 was used as a strategy to identify genes required for synthesis of the Galα(1–4)βGal component of Haemophilus influenzae strain RM7004 lipopolysaccharide. Insertion of Tn916 into an open reading frame (ORF) encoding a protein with 75% homology to the Escherichia coli methionine related protein (Mrp) is described. Mutations in mrp resulted in loss of reactivity with monoclonal antibody (mAb) 4C4, which recognises Galα(1–4)βGal, and expression of LPS with a different electrophoretic profile to that of wild‐type RM7004. An unexpected feature of this mutation was that it appeared to influence the number of copies of 5′‐CAAT‐3′ present in lic2A , a gene which is also required for biosynthesis and phase variable expression of the Galα(1–4)βGal LPS epitope.