Identification of rfbA, involved in B-band lipopolysaccharide biosynthesis in Pseudomonas aeruginosa serotype O5.

Infection and Immunity
T Dasgupta, J S Lam

Abstract

Previous work from this laboratory has shown that a 26-kb insert in cosmid clone pFV100, isolated from a Pseudomonas aeruginosa gene library, contained genes that could restore serotype-specific B-band lipopolysaccharide (LPS) expression in rough mutant ge6. In this study, subclones from pFV100 were made to identify genes responsible for B-band LPS synthesis. Transformation of Escherichia coli HB101 with cosmid clone pFV100 resulted in expression of P. aeruginosa serotype O5 B-band LPS, indicating the presence of an rfb cluster in pFV100. Expression of P. aeruginosa LPS could not be achieved in E. coli HB101 transformed with any of the subclones. Complementation studies of well-characterized rough mutants of P. aeruginosa PAO1 deficient in B-band LPS biosynthesis were performed with the various subclones. Subclone pFV110, containing a 1.4-kb XbaI-HindIII insert, restored B-band LPS biosynthesis in mutant AK44 (A+B-; complete core). Probing chromosomal DNA from the 20 International Antigenic Typing Scheme serotypes with the 1.4-kb insert from pFV110 in Southern hybridizations revealed a positive reaction to restriction fragments in serotypes O2, O5, O16, O20, and O18. LPS of serotypes O2, O5, O16, and O20 were shown earlier to h...Continue Reading

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Citations

Mar 27, 2015·Evolution; International Journal of Organic Evolution·Sean MeadenBritt Koskella
Jul 10, 1999·Journal of Bacteriology·C R DeanJ B Goldberg
Aug 26, 2020·Journal of Bacteriology·Sara M KleeTimothy W McNellis
Dec 1, 1996·Trends in Microbiology·J B Goldberg, G B Pler
Sep 8, 1999·Microbiology and Molecular Biology Reviews : MMBR·H L RocchettaJ S Lam
Dec 1, 1995·Infection and Immunity·G B Pier, J B Goldberg

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