Isolation of Zoogloea ramigera I-16-M exopolysaccharide biosynthetic genes and evidence for instability within this region.

TitleIsolation of Zoogloea ramigera I-16-M exopolysaccharide biosynthetic genes and evidence for instability within this region.
Publication TypeJournal Article
Year of Publication1987
AuthorsEasson, DD, Sinskey, AJ, Peoples, OP
JournalJ Bacteriol
Volume169
Issue10
Pagination4518-24
Date Published1987 Oct
ISSN0021-9193
KeywordsChromosome Mapping, Cloning, Molecular, Conjugation, Genetic, DNA Restriction Enzymes, DNA Transposable Elements, DNA, Bacterial, Genes, Bacterial, Genetic Complementation Test, Mutation, Nucleic Acid Hybridization, Polysaccharides, Bacterial, Zoogloea
Abstract

The genetics of the biosynthesis of an exocellular polysaccharide (EPS) from Zoogloea ramigera I-16-M is being investigated. Tn5 insertion mutants deficient in EPS production were isolated by screening for the absence of fluorescence on plates containing the dye Cellufluor (Polysciences Chemicals). Complementation of these mutations was achieved with a Z. ramigera I-16-M gene library constructed in a broad-host-range cosmid vector and introduced into the I-16-M mutants by conjugation. Four recombinant plasmids able to restore EPS production to all of these mutants were found to contain at least 14 kilobases of common insert DNA. Subcloning of the common region and restriction mapping the locations of Tn5 insertions have identified two complementation groups contained within a chromosomal segment of DNA that is between 4.6 and 6.5 kilobases in size. We have clearly demonstrated genetic instability in this region which leads to spontaneous deletions and possibly rearrangements resulting in the loss of EPS production.

DOI10.1128/jb.169.10.4518-4524.1987
Alternate JournalJ Bacteriol
Citation Key202
PubMed ID2820929
PubMed Central IDPMC213816