Search for the atp Operon URF6 Gene in Rhodobacter sphaeroides and Paracoccus denitrificans and Partial Sequencing of the Corresponding atpD and atpG Genes (original) (raw)

1998

Abstract

The F-type ATP synthase supplies the cell with ATP using the energy from proton gradients across membranes to phosphorylate ADP. The F1 catalytic subunit consists of five subunits: α, β, γ, δ and e. The genes encoding these subunits in bacteria are clustered in one operon, called the atp operon. The conserved gene order in the atp operons of chloroplasts, Escherichia coli, Rhodospirillum rubrum, Rhodobacter blasticus and some cyanobacteria, indicates common ancestry [1]. An interesting feature of the Rb. blasticus atp operon is that a sixth gene, termed URF6, is placed between the genes for the γ and β subunits [2]. Since the genes of the atp operon are co-transcribed, the URF6 gene product may be involved in the ATP synthase. Previously, we have raised antibodies towards the URF6 gene product, and used them to investigate the presence of this protein in several other bacteria and during purification of the overexpressed recombinant protein [3]. Our studies indicate that Paracoccus denitrificans also appears to contain this protein. In the present study we have used a PCR-based method and Southern blotting to investigate if the URF6 gene is present in P. denitrificans and Rhodobacter sphaeroides and to determine its location in the genome. The PCR strategy used primers designed from conserved regions at the end of the atpG gene (γ) and beginning of the atpD gene (β). PCR products with the expected sizes were cloned and sequenced.

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