Cloning and expression of draTG genes from Azospirillum lipoferum.
A genomic library of Azospirillum lipoferum was constructed with phage lambda EMBL4 as vector. From this library, the genes encoding dinitrogenase reductase ADP-ribosyltransferase (DRAT), draT, and dinitrogenase reductase-activating glycohydrolase (DRAG), draG, were cloned by hybridization with the heterologous probes of Rhodospirillum rubrum. As in R. rubrum, draT is located between draG and nifH, the gene encoding dinitrogenase reductase (a substrate for the DRAG/DRAT system). In the crude extract of Escherichia coli harboring the expression vector for this region, DRAT and DRAG enzyme activities were detected, confirming the identity of the cloned genes. Southern hybridization with genomic DNA from different Azospirillum spp., demonstrated a correlation between observable draTG hybridization and the biochemical demonstration of this covalent modification system.
Fu
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Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin-Madison 53706.
Fitzmaurice
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Roberts
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Burris
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D036002:ADP Ribose Transferases / Q000235:genetics*; D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D006026:Glycoside Hydrolases / Q000235:genetics*; D006090:Gram-Negative Bacteria / Q000235:genetics*; D009699:N-Glycosyl Hydrolases; D009586:Nitrogen Fixation / Q000235:genetics*; D015183:Restriction Mapping
D004269:DNA, Bacterial; D000246:Adenosine Diphosphate Ribose; D036002:ADP Ribose Transferases; C057637:dinitrogenase reductase ADP-ribosyltransferase; D006026:Glycoside Hydrolases; D009699:N-Glycosyl Hydrolases; C036091:ADP-ribosyl-(dinitrogen reductase) hydrolase