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PMID: 1657881 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Cloning, characterization, and DNA sequence of the rfaLK region for lipopolysaccharide synthesis in Salmonella typhimurium LT2.

Journal of bacteriology ·Vol. 173 ·No. 22 ·1991-11-00 ·Pages 7151-63

MacLachlan PR, Kadam SK, Sanderson KE

Abstract

We have cloned and sequenced the rfaL and rfaK genes for lipopolysaccharide synthesis in Salmonella typhimurium LT2 on a 4.28-kb HindIII fragment from the previously described R' factor pKZ3 (S. K. Kadam, A. Rehemtulla, and K. E. Sanderson, J. Bacteriol. 161:277-284, 1985). rfaL is thought to encode a component of the O-antigen ligase, and rfaK is believed to encode the N-acetylglucosamine transferase. The genes were identified by the loss of complementation of prototype rfaL and rfaK mutations after Tn1000 mutagenesis. Translation of the nucleotide sequence predicted sizes of 45.9 and 43.1 kDa for the rfaL and rfaK gene products, respectively. Hydropathy analysis of the rfaL product suggested that it was an integral membrane protein. A third gene, rfaZ, was found to be an 808-bp open reading frame on the pyrE side of rfaK. Insertions into rfaZ reduced rfaK complementation, suggesting cotranscription in the pyrE-cysE direction. The rfaL gene is transcribed in the opposite direction in a separate operon which may also include rfaC. An incomplete open reading frame with homology to an Escherichia coli gene in the same region, rfaY, was found on the pyrE side of rfaZ. Complementation studies with Tn1000 insertions in rfaL showed that rfaL446 and rfaL447 are allelic. With the cloning of the rfaL and -K genes, the order of genes within the rfa cluster at 79 units on the linkage map was found to be cysE-rfaDFCLKZYJIBG-pyrE.

Related Genes
rfaC rfaK rfaL rfaZ
MeSH Terms
Amino Acid Sequence Base Sequence Blotting, Southern Carbohydrate Sequence Cloning, Molecular/methods DNA Transposable Elements DNA, Bacterial/genetics Escherichia coli/genetics Genes, Bacterial Genetic Complementation Test Genotype Lipopolysaccharides/biosynthesis Molecular Sequence Data Mutagenesis, Insertional Open Reading Frames Operon Protein Conformation Restriction Mapping Salmonella typhimurium/enzymology,genetics Sequence Homology, Nucleic Acid
Chemicals
DNA Transposable Elements DNA, Bacterial Lipopolysaccharides
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
MacLachlan P R
Department of Biological Sciences, University of Calgary, Alberta, Canada.
Kadam S K
Sanderson K E
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.173.22.7151-7163.1991
Published
1991-11-00
Pages
7151-63
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC209221
Subset
IM
Databases
GENBANK
M63251, M63252, M63253, M63254, M63255, M73826, S59274, S60896, S65089, S65094
External Links
PubMed source DOI: 10.1128/jb.173.22.7151-7163.1991 PMC Full Text View journal details
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