Cloning of aroG, the gene coding for phospho-2-keto-3-deoxy-heptonate aldolase(phe), in Escherichia coli K-12, and subcloning of the aroG promoter and operator in a promoter-detecting plasmid.
Defective transducing phages carrying aroG, the structural gene for phenylalanine (phe)-inhibitable phospho-2-keto-heptonate aldolase (EC 4.1.2.15; previously known as 3-deoxy-D-arabinoheptulosonate-7-phosphate synthetase[phe]), have been isolated, and DNA from two of these phages has been used to construct a restriction map of the region from att lambda to aroG. A 7.6-kb PstI-HindIII fragment from one of these phages was cloned into pBR322 and shown to contain aroG. The location of aroG within the 7.6 kb was established by subcloning and Tn3 transpositional mutagenesis. A fragment carrying the aroG promoter and operator has been cloned into a high copy number promoter-cloning vector (pMC489), and the resulting aroGpo-LacZ' (alpha) fusion subcloned in a low copy number vector. Strains with this fusion on the low copy number vector exhibit negative regulation of beta-galactosidase expression by both phenylalanine and tryptophan and positive regulation by tyrosine in a tyrR+ background.
Davies
W D
WD
Pittard
J
J
Davidson
B E
BE
D010723:3-Deoxy-7-Phosphoheptulonate Synthase / Q000235:genetics*; D000446:Aldehyde-Lyases / Q000235:genetics*; D010582:Bacteriophage lambda / Q000235:genetics; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D009154:Mutation; D010957:Plasmids; D011401:Promoter Regions, Genetic; D001616:beta-Galactosidase / Q000235:genetics
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004279:DNA, Viral; D010723:3-Deoxy-7-Phosphoheptulonate Synthase; D004262:DNA Restriction Enzymes; D001616:beta-Galactosidase; D000446:Aldehyde-Lyases