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PMID: 6086327 Published · ppublish English Journal Article

Molecular cloning, sequencing, and expression of the crr gene: the structural gene for IIIGlc of the bacterial PEP:glucose phosphotransferase system.

The EMBO journal ·Vol. 3 ·No. 7 ·1984-07-00 ·Pages 1587-93

Nelson SO, Schuitema AR, Benne R, van der Ploeg LH, Plijter JS, Aan F, Postma PW

Abstract

The phosphoenolpyruvate:glucose phosphotransferase system (PTS) of Salmonella typhimurium is involved both in glucose transport and in the regulation and synthesis of adenylate cyclase and several transport systems. The crr gene has been implicated in this regulating mechanism. A 9.6-kb segment of the S. typhimurium chromosome containing the crr gene was cloned in pAT153. The cloned fragment also complemented cysA mutations but did not contain a functional pts operon which is closely linked to the crr gene and codes for two enzymes of the PTS. Although cysA and crr have been reported to be located on opposite sides of ptsHI, our results suggest that the correct gene order is cysK-ptsHI-crr-cysA. Expression of crr plasmids in a maxicell system yielded two proteins which reacted with specific anti-serum against IIIGlc. The apparent mol. wts. in SDS-polyacrylamide gels were 20 000 and 21 000, the former corresponding to the major band of purified IIIGlc. Both forms were also observed in bacterial extracts and purified IIIGlc. The crr gene was localized on a 1-kb EcoRI-EcoRV fragment of the 9.6-kb insert and sequenced. It codes for a single protein (18 556 D) containing 169 amino acid residues and identified as IIIGlc.

MeSH Terms
Base Sequence Chromosome Mapping Cloning, Molecular DNA Restriction Enzymes Escherichia coli/genetics Gene Amplification Gene Expression Regulation Genes Genes, Bacterial Macromolecular Substances Phosphoenolpyruvate Sugar Phosphotransferase System/genetics Salmonella typhimurium/genetics
Chemicals
Macromolecular Substances Phosphoenolpyruvate Sugar Phosphotransferase System phosphoenolpyruvate-glucose phosphotransferase DNA Restriction Enzymes
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Nelson S O
Schuitema A R
Benne R
van der Ploeg L H
Plijter J S
Aan F
Postma P W
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1984-07-00
Pages
1587-93
Language
English
Region
England
NLM ID
8208664
PMCID
PMC557563
Subset
IM
Databases
GENBANK
X05210
External Links
PubMed source PMC Full Text View journal details
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