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J Bacteriol, June 1998, p. 3056-3061, Vol. 180, No. 12
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Replicase, Excisionase, and Integrase Genes of the Streptomyces Element pSAM2 Constitute an Operon Positively Regulated by the pra Gene

Guennadi Sezonov,* Anne-Marie Duchêne,dagger Annick Friedmann, Michel Guérineau, and Jean-Luc Pernodet

Laboratoire de Biologie et Génétique Moléculaire, Institut de Génétique et Microbiologie, URA CNRS 2225, Université Paris-Sud, 91405 Orsay, France

Received 24 November 1997/Accepted 13 April 1998

pSAM2 is a site-specific integrative element from Streptomyces ambofaciens. The pra gene described earlier as an activator of pSAM2 replication is shown here to be also involved in the activation of its integration and excision. This was evidenced with derivatives of pSAM2 mutant B3 in which the pra gene was placed under the control of the inducible tipAp promoter. Transformation of Streptomyces lividans by these derivatives was efficient only when pra expression was induced, indicating its involvement in pSAM2 integration activation. Once established, these constructions remained integrated in the chromosome under noninduced conditions. Activation of the pra expression provoked strong activation of their excision, leading to the appearance of free forms. The results of functional, transcriptional, and sequence analyses allowed to conclude that the three genes repSA, xis, and int coding for the pSAM2 replicase, excisionase, and integrase, respectively, constitute an operon directly or indirectly activated by pra.

* Corresponding author. Mailing address: Laboratoire de Biologie et Génétique Moléculaire, Institut de Génétique et Microbiologie, URA CNRS 2225, Bâtiment 400, Université Paris-Sud, 91405 Orsay, France. Phone: 33-(0)-1-69-15-46-40. Fax: 33-(0)-1-69-15-72-96. E-mail: sezonov{at}igmors.u-psud.fr.

dagger Present address: Institut de Biologie Moléculaire des Plantes, 67084 Strasbourg, France.

J Bacteriol, June 1998, p. 3056-3061, Vol. 180, No. 12
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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