SUMMARY: We have previously isolated ineffective (Fix) mutants of 104A14 requiring both arginine and uracil, and thus probably defective in carbamoylphosphate synthetase. We describe here the molecular and genetic analysis of the genes coding for carbamoylphosphate synthetase. Plasmids that complement the mutations were isolated from a gene bank. Restriction analysis of these plasmids indicated that complementation involved two unlinked regions of the chromosome, and Genetic complementation between the plasmids and mutants demonstrated a single complementation group for , but two overlapping complementation groups for The cloned genes hybridize to the corresponding and genes which encode the two subunits of carbamoylphosphate synthetase. Transposon Tn5 mutagenesis was used to localize the and genes on the cloned DNA. The cloned and genes were unable to complement or mutants alone or in combination. We speculate on the mechanism of the unusual pattern of genetic complementation at the locus.


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