Cloning, nucleotide sequence, and mutagenesis of a gene (irpA) involved in iron-deficient growth of the cyanobacterium Synechococcus sp. strain PCC7942

Journal Article · Sat Oct 01 04:00:00 EDT 1988 · Journal of Bacteriology; (USA)

OSTI ID:7067534

Reddy, K J; Bullerjahn, G S; Sherman, D M; Sherman, L A ^[1]

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1. Univ. of Missouri, Columbia (USA)

We describe the cloning and sequencing of a gene from the cyanobacterium Synechococcus sp. strain PCC7942, designated irpA (iron-regulated protein A), that encodes for a protein involved in iron acquisition or storage. Polyclonal antibodies raised against proteins which accumulate during iron-deficient growth were used as probes to isolate immunopositive clones from a {lambda}gt11 genomic expression library. The clone, designated {lambda}gtAN26, carried a 1.7-kilobase (kb) chromosomal DNA insert and was detected by cross-reactivity with antibody against a 36-kilodalton protein. It was possible to map a 20-kb portion of the chromosome with various DNA probes from {lambda}gt11 and {lambda}EMBL-3 clones, and Southern blot analysis revealed that the irpA gene was present in a single copy and localized within a 1.7-kb PstI fragment. DNA sequencing revealed an open reading frame of 1,068 nucleotides capable of encoding 356 amino acids which yields a protein with a molecular weight of 38,584. The hydropathy profile of the polypeptide indicated a putative N-terminal signal sequence of 44 amino acid residues. IrpA is a cytoplasmic membrane protein as determined by biochemistry and electron microscopy immunocytochemistry. The upstream region of the irpA gene contained a consensus sequence similar to the aerobactin operator in Escherichia coli.

• DOE Contract Number: FG02-86ER13516
• OSTI ID: 7067534
• Journal Information: Journal of Bacteriology; (USA), Journal Name: Journal of Bacteriology; (USA) Vol. 170:10; ISSN JOBAA; ISSN 0021-9193
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
BIOLOGICAL AVAILABILITY
CLONING
CYANOBACTERIA
DNA HYBRIDIZATION
DNA SEQUENCING
DNA-CLONING
ELEMENTS
GENE REGULATION
HYBRIDIZATION
IRON
METALS
MICROORGANISMS
MOLECULAR BIOLOGY
MUTAGENESIS
ORGANIC COMPOUNDS
PROTEINS
STRUCTURAL CHEMICAL ANALYSIS
TRANSITION ELEMENTS