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PMID: 2549367 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Cloning of a gene involved in cellulose biosynthesis in Acetobacter xylinum: complementation of cellulose-negative mutants by the UDPG pyrophosphorylase structural gene.

Molecular & general genetics : MGG ·Vol. 217 ·No. 1 ·1989-05-00 ·Pages 26-30

Valla S, Coucheron DH, Fjaervik E, Kjosbakken J, Weinhouse H, Ross P, Amikam D, Benziman M

Abstract

Three cellulose-negative (Cel-) mutants of Acetobacter xylinum strain ATCC 23768 were complemented by a cloned 2.8 kb DNA fragment from the wild type. Biochemical analysis of the mutants showed that they were deficient in the enzyme uridine 5'-diphosphoglucose (UDPG) pyrophosphorylase. The analysis also showed that the mutants could synthesize beta(1-4)-glucan in vitro from UDPG, but not in vivo from glucose. This result was expected, since UDPG is known to be the precursor for cellulose synthesis in A. xylinum. In order to analyze the function of the cloned gene in more detail, its biological activity in Escherichia coli was studied. These experiments showed that the cloned fragment could be used to complement an E. coli mutant deficient in the structural gene for UDPG pyrophosphorylase. It is therefore clear that the cloned fragment must contain this gene from A. xylinum. This is to our knowledge the first example of the cloning of a gene with a known function in cellulose biosynthesis from any organism, and we suggest the gene be designated celA.

MeSH Terms
Cellulose/biosynthesis,genetics Cloning, Molecular Conjugation, Genetic DNA, Bacterial/genetics Electrophoresis, Agar Gel Escherichia coli Proteins Genes Genetic Complementation Test Gluconacetobacter xylinus/enzymology,genetics Lyases/metabolism Mutation Nucleotidyltransferases/genetics Phosphoglucomutase/metabolism Phosphoric Diester Hydrolases/metabolism Phosphorus-Oxygen Lyases Plasmids UTP-Glucose-1-Phosphate Uridylyltransferase/genetics
Chemicals
DNA, Bacterial Escherichia coli Proteins Cellulose Nucleotidyltransferases UTP-Glucose-1-Phosphate Uridylyltransferase Phosphoric Diester Hydrolases Lyases Phosphorus-Oxygen Lyases diguanylate cyclase Phosphoglucomutase
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Valla S
Laboratory of Biotechnology, Norwegian Institute of Technology, Trondheim.
Coucheron D H
Fjaervik E
Kjosbakken J
Weinhouse H
Ross P
Amikam D
Benziman M
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00330938
Published
1989-05-00
Pages
26-30
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00330938
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