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Disruption of sblA in Streptomyces lividans permits expression of a heterologous -amylase gene in the presence of glucose

Laboratoire de Biologie et Génétique Moléculaire, Institut de Génétique et Microbiologie, CNRS UMR8621 Bâtiment 400, Université Paris-Sud, F-91405 Orsay Cedex, France¹

Author for correspondence: Marie-Joelle Virolle. Tel: +33 169156913. Fax: +33 169156678. e-mail: virolle{at}igmors.u-psud.fr

In a transposition mutant of Streptomyces lividans TK24, the usually glucose-repressible expression of a heterologous -amylase gene (aml) became resistant to glucose repression. The transposon had inserted into an ORF called sblA which encodes a 274 aa product sharing significant sequence similarities with various phosphatases that act on small phosphorylated substrates. sblA was transcribed as a monocistronic mRNA and its transcription was enhanced at the transition phase. Because its transcriptional and putative translational start points coincide, sblA is likely to be translated in the absence of a conventional RBS. The sblA-disrupted mutant is characterized by early growth arrest in glucose-grown cultures and by partial relief of glucose repression of aml expression.

Keywords: glucose repression, induction, inositol monophosphatase, transition phase

Abbreviations: Amp, ampicillin; Apr, apramycin; CBS, Cibachron blue starch; IMP, inositol monophosphatase; Nos, nosiheptide

The EMBL accession number for the sequence reported in this paper is AJ223365.

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