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A 15437 bp region encompassing the inhA locus from the Mycobacterium awium chromosome was cloned and sequenced. From the sequencing data generated and the results of homology searches, the primary structure of this region was determined. This region contains four known genes (acnA, fabG, inhA and hemH) and two genes, inwA and inwB, whose products display homology with p60 invasion protein of Listeria monocytogenes. Six proteins encoded by putative ORFs contained an RGD motif (often involved in binding to macrophage integrins), while ORFl and MoxR are probably transcriptional regulators. The rest of the putative products encoded by ORFs in the sequenced region showed little homology with the proteins contained in the databases and were considered to be unknown proteins.

The product of the MTCY428.08 gene ofMycobacterium tuberculosis shows sequence homology with several NAD+ synthetases. The MTCY428.08 gene was cloned into the expression vectors pGEX-4T-1 and pET-15b. Expression in Escherichia coli led to overproduction of glutathione S-transferase fused and His6-tagged gene products, which were enzymatically assayed for NAD synthetase activity. Our results demonstrate that the MTCY428.08 gene of M. tuberculosis is the structural gene for NAD+ synthetase.

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