Molecular Cloning and Expression of the IlvGEDAY Genes from Salmonella Typhimurium

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1981 Aug 1

PMID 6167564

Citations 12

Authors

D L Blazey

R Kim

R O BURNS

Affiliations

Soon will be listed here.

Abstract

The ilvGEDAY genes of Salmonella typhimurium were cloned in Escherichia coli K-12 by in vitro recombination techniques. A single species of recombinant plasmid, designated pDU1, was obtained by selecting for Valr Ampr transformants of strain SK1592. pDU1 was shown to contain a 14-kilobase EcoRI partial digestion product of the S. typhimurium chromosome inserted into the EcoRI site of the pVH2124 cloning vector. The ilvGEDAY genes were found to occupy a maximum length of 7.5 kilobases. Restriction endonuclease analysis of the S. typhimurium ilv gene cluster provided another demonstration of the gene order as well as established the location of ilv Y between ilvA and ilvC. The presence of a ribosomal ribonucleic acid operon on the pDU1 insert, about 3 kilobases from the 5' end of ilvG, was shown by Southern hybridization. The expression of the ilvGEDA operon from pDU1 was found to be elevated, reflecting the increased gene dosage of the multicopy plasmid. A polarity was observed with respect to ilvEDA expression which is discussed in terms of the possible translational effects of the two internal promoter sequences, one located proximal to ilvE and the other located proximal to ilvD.

Citing Articles

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L-alanine auxotrophy of Lactobacillus johnsonii as demonstrated by physiological, genomic, and gene complementation approaches.

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Altered regulation of isoleucine-valine biosynthesis in a hisW mutant of Salmonella typhimurium.

Davis L, Williams L J Bacteriol. 1982; 151(2):860-6.

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Metabolic basis for the isoleucine, pantothenate or methionine requirement of ilvG strains of Salmonella typhimurium.

Primerano D, BURNS R J Bacteriol. 1982; 150(3):1202-11.

PMID: 7042686 PMC: 216341. DOI: 10.1128/jb.150.3.1202-1211.1982.

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