1887

Abstract

Amplification of sequences from ISP5230 genomic DNA using PCR with primers based on conserved prokaryotic sequences gave two main products. One matched , a locus previously identified in The second closely resembled the conserved sequence consensus and hybridized with a 38 kb I fragment of ISP5230 genomic DNA. Cloning and sequence analysis of the 38 kb fragment detected three ORFs, and their deduced amino acid sequences were used in BLAST searches of the GenBank database. The ORF1 product was similar to PabB in other bacteria and to the PabB domain encoded by . The ORF2 product resembled PabA of other bacteria. ORF3 was incomplete; its deduced partial amino acid sequence placed it in the MocR group of GntR-type transcriptional regulators. Introducing vectors containing the 38 kb I fragment of DNA into and mutants of , or into the mutant JG10, enhanced sulfanilamide resistance in the host strains. The increased resistance was attributed to expression of the pair of discrete translationally coupled -aminobenzoic acid biosynthesis genes (designated /) cloned in the 38 kb fragment. These represent a second set of genes encoding 4-amino-4-deoxychorismate synthase in ISP5230. In contrast to the fused set previously isolated from this species, they do not participate in chloramphenicol biosynthesis, but like they can be disrupted without affecting growth on minimal medium. The gene disruption results suggest that may have a third set of genes encoding PABA synthase.

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2001-08-01
2020-01-20
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