and are pathogens responsible for disease in both humans and animals. , while phylogenetically similar, is considered avirulent in comparison. These three species exhibit phosphatase activity when grown on media containing chromogenic substrates such as 5-bromo-4-chloro-3-indolyl phosphate (XP). Tn-OT182 mutagenesis has been utilized to isolate mutants of and unable to hydrolyse XP. Sequence analysis of these mutants revealed an ORF of 1734 nucleotides demonstrating a high degree of homology to the gene product of . PCR primers were designed based on the gene sequence and were used to amplify an homologue from . The predicted amino acid sequence of AcpA differed from those of the predicted AcpA and AcpA by 15 and 3 amino acids, respectively. Allelic exchange was used to construct Δ mutants in each of these spp. These mutants were shown to be devoid of phosphatase activity and have subsequently allowed for the implementation of fusion transposon mutagenesis systems. Two such systems have been successfully utilized in spp. for the identification of several genes encoding exported proteins.


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