Genes from the purine biosynthetic pathway were identified using purine auxotrophic mutants of obtained by Tn transposon mutagenesis. Two approaches were followed in parallel. The first consisted of the complementation of the purine auxotrophs using a H37Rv shuttle cosmid library. In the second approach, specific probes corresponding to the regions adjacent to the insertion sites of Tn in the genome were used to screen a plasmid library by colony hybridization for inserts carrying homologous DNA fragments. Nucleotide sequence analysis of two genes isolated by these methods revealed high similarities with and genes from other bacterial and fungal sources. Transcriptional start sites were mapped for both genes, which revealed similar −10 boxes but with a higher GC content than the σ consensus.


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