PMID:1711024
| Citation |
Pi, J, Wookey, PJ and Pittard, AJ (1991) Cloning and sequencing of the pheP gene, which encodes the phenylalanine-specific transport system of Escherichia coli. J. Bacteriol. 173:3622-9 |
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| Abstract |
The phenylalanine-specific permease gene (pheP) of Escherichia coli has been cloned and sequenced. The gene was isolated on a 6-kb Sau3AI fragment from a chromosomal library, and its presence was verified by complementation of a mutant lacking the functional phenylalanine-specific permease. Subcloning from this fragment localized the pheP gene on a 2.7-kb HindIII-HindII fragment. The nucleotide sequence of this 2.7-kb region was determined. An open reading frame was identified which extends from a putative start point of translation (GTG at position 636) to a termination signal (TAA at position 2010). The assignment of the GTG as the initiation codon was verified by site-directed mutagenesis of the initiation codon and by introducing a chain termination mutation into the pheP-lacZ fusion construct. A single initiation site of transcription 30 bp upstream of the start point of translation was identified by the primer extension analysis. The pheP structural gene consists of 1,374 nucleotides specifying a protein of 458 amino acid residues. The PheP protein is very hydrophobic (71% nonpolar residues). A topological model predicted from the sequence analysis defines 12 transmembrane segments. This protein is highly homologous with the AroP (general aromatic transport) system of E. coli (59.6% identity) and to a lesser extent with the yeast permeases CAN1 (arginine), PUT4 (proline), and HIP1 (histidine) of Saccharomyces cerevisiae. |
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| Keywords |
Amino Acid Sequence; Amino Acid Transport Systems; Amino Acid Transport Systems, Neutral; Bacterial Proteins/chemistry; Bacterial Proteins/genetics; Base Sequence; Biological Transport; Carrier Proteins/chemistry; Chromosomes, Bacterial; Cloning, Molecular; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli Proteins; Genes, Bacterial; Membrane Transport Proteins/genetics; Molecular Sequence Data; Nucleic Acid Hybridization; Open Reading Frames; Phenylalanine/metabolism; Protein Biosynthesis; Protein Conformation; RNA, Bacterial/genetics; Restriction Mapping; Sequence Homology, Nucleic Acid; Transcription, Genetic |
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