Applied Genetics and Molecular Biotechnology

Applied Microbiology and Biotechnology

, Volume 68, Issue 5, pp 656-662

Expression in Streptomyces lividans of Nonomuraea genes cloned in an artificial chromosome

  • Rosa AlduinaAffiliated withDipartimento Biologia Cellulare e dello Sviluppo, Viale delle Scienze, University of Palermo Email author 
  • , Anna GiardinaAffiliated withDipartimento Biologia Cellulare e dello Sviluppo, Viale delle Scienze, University of Palermo
  • , Giuseppe GalloAffiliated withDipartimento Biologia Cellulare e dello Sviluppo, Viale delle Scienze, University of Palermo
  • , Giovanni RenzoneAffiliated withProteomics and Mass Spectrometry laboratory, ISPAAM
  • , Clelia FerraroAffiliated withDipartimento Biologia Cellulare e dello Sviluppo, Viale delle Scienze, University of Palermo
  • , Alba ContinoAffiliated withDipartimento Biologia Cellulare e dello Sviluppo, Viale delle Scienze, University of Palermo
  • , Andrea ScaloniAffiliated withProteomics and Mass Spectrometry laboratory, ISPAAM
  • , Stefano DonadioAffiliated withVicuron Pharmaceuticals, Microbial Technologies
  • , Anna Maria PugliaAffiliated withDipartimento Biologia Cellulare e dello Sviluppo, Viale delle Scienze, University of Palermo

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Abstract

A bacterial artificial chromosomal library of Nonomuraea sp. ATCC39727 was constructed using Escherichia coliStreptomyces artificial chromosome (ESAC) and screened for the presence of dbv genes known to be involved in the biosynthesis of the glycopeptide A40926. dbv genes were cloned as two large, partially overlapping, fragments and transferred into the host Streptomyces lividans, thus generating strains S. lividans∷NmESAC50 and S. lividans∷NmESAC57. The heterologous expression of Nonomuraea genes in S. lividans was successfully demonstrated by using combined RT–PCR and proteomic approaches. MALDI-TOF analysis revealed that a Nonomuraea ABC transporter is expressed as two isoforms in S. lividans. Moreover, its expression may not require a Nonomuraea positive regulator at all, as it is present at similar levels in both clones even though S. lividans∷NmESAC57 lacks regulatory genes. Considered together, these results show that S. lividans expresses Nonomuraea genes from their own promoters and support the idea that S. lividans can be a good host for genetic analysis of Nonomuraea.