Cloning, nucleotide sequence, and mutagenesis of a gene (irpA) involved in iron-deficient growth of the cyanobacterium Synechococcus sp. strain PCC7942
- 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), Vol. 170:10; ISSN 0021-9193
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
CYANOBACTERIA
MOLECULAR BIOLOGY
IRON
BIOLOGICAL AVAILABILITY
PROTEINS
DNA SEQUENCING
DNA-CLONING
GENE REGULATION
MUTAGENESIS
CLONING
DNA HYBRIDIZATION
ELEMENTS
HYBRIDIZATION
METALS
MICROORGANISMS
ORGANIC COMPOUNDS
STRUCTURAL CHEMICAL ANALYSIS
TRANSITION ELEMENTS
550200* - Biochemistry