An agar plating technique was developed in which the activation of expressior of a gene fusion was used to isolate the genes from a gene library. An mutant containing the fusion was transformed and plated on a low-nitrogen medium so that on flooding with ONPG, the production of yellow colonies indicated the presence of the cloned genes. A 4.47 kb region from the chromosome was sequenced. Analysis of the sequence revealed that the and genes were closely linked to a third ORF of unknown function. Analysis of the 900 bp region upstream of the genes and Southern hybridization experiments confirmed that in ATCC 33020, the and genes are unlinked. Expression of the fusion in was activated in the presence of the genes and regulated by nitrogen.


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