Cloning of a mineral phosphate-solubilizing gene from XXXX .

@entity6402 @entity535 @entity3793 @entity31938 @entity377 @entity460 @entity12742 @entity413 @entity2624 @entity526 @entity31937
We have recently shown that the ability of some gram-negative bacteria to dissolve poorly soluble @entity460 phosphates (Mps+ phenotype) is the result of periplasmic oxidation of @entity413 to @entity12742 via the quinoprotein @entity413 dehydrogenase (GDH), a component of the direct oxidation pathway. @entity2624 derivatives synthesize apo-GDH but not the cofactor @entity31937 ( @entity31938 ) essential for formation of the holoenzyme. Therefore, in the absence of exogenous @entity31938 , these strains do not produce @entity12742 and are Mps-. Evidence is presented to show that expression of a single 396-base @entity6402 open reading frame (designated gabY) in @entity526 JM109 (a @entity2624 derivative) was sufficient to induce the Mps+ phenotype and production of @entity12742 . We present the @entity377 sequence of this open reading frame which coded for a protein (GabY) with a deduced M(r) of 14,235. Coupled transcription-translation of a plasmid (pSLY4 or pGAB1) carrying gabY resulted in production of a protein with an M(r) of 14,750. Disruption of the open reading frame of gabY via site-directed mutagenesis changed the phenotype to Mps- and eliminated @entity12742 production. The deduced @entity535 sequence of gabY has no apparent homology with those of previously cloned direct oxidation pathway genes but does share regions highly homologous with the @entity3793 permease system membrane-bound protein HisQ as well as other proteins in this family. In the presence of 1 microM exogenous @entity31938 , both JM109(pSLY4) and JM109(pGAB1) produced 10 times as much @entity12742 as was seen with either the plasmid or exogenous @entity31938 alone.(ABSTRACT TRUNCATED AT 250 WORDS)
