AraL sugar phosphatase from B. subtilis L. M. Godinho and I. de Sa´-Nogueira 34 De´ barbouille´ M, Arnaud M, Fouet A, Klier A & Rapo- port G (1990) The sacT gene regulating the sacPA operon in Bacillus subtilis shares strong homology with transcriptional antiterminators. J Bacteriol 172, 3966– 3973. 35 Sambrook J, Fritsch EF & Maniatis T (1989) Molecular Cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. 36 Huggins C & Smith DR (1947) Chromogenic substrates III. p-nitrophenyl sulfate as a substrate for the assay of phenolsulfatase activity. J Biol Chem 170, 391–398. 37 Arnaud M, Chastanet A & D e´barbouille´ M (2004) New vector for efﬁcient allelic replacement in naturally non- transformable, low GC content, gram-positive bacteria. Appl Environ Microbiol 70, 6887–6891. 38 Anagnostoupoulos C & Spizizen J (1961) Requirements for transformation in Bacillus subtilis. J Bacteriol 81, 741–746. 39 Weinrauch Y, Msadek T, Kunst F & Dubnau D (1991) Sequence and properties of comQ, a new competence regulatory gene of Bacillus subtilis. J Bacteriol 173, 5685–5693. 40 Studier FW, Rosenberg AH, Dunn JJ & Dubendorff JW (1990) Use of T7 RNA polymerase to direct expres- sion of cloned genes. Methods Enzymol 185, 60–89. 41 Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R et al. (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23, 2947–2948. 2524 FEBS Journal 278 (2011) 2511–2524 ª 2011 The Authors Journal compilation ª 2011 FEBS