title: Expression in Streptomyces lividans of Nonomuraea genes cloned in an artificial chromosome.
abstract: A bacterial artificial chromosomal library of Nonomuraea sp. ATCC39727 was constructed using Escherichia coli-Streptomyces 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. lividansColon, two colonsNmESAC50 and S. lividansColon, two colonsNmESAC57. 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. lividansColon, two colonsNmESAC57 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.
authors.affiliation: Dipartimento Biologia Cellulare e dello Sviluppo, Viale delle Scienze, University of Palermo, Parco d'Orleans II, Italy. valduina@unipa.it
authors.forename: Rosa
authors.initials: R
authors.lastname: Alduina
authors.forename: Anna
authors.initials: A
authors.lastname: Giardina
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Gallo
authors.forename: Giovanni
authors.initials: G
authors.lastname: Renzone
authors.forename: Clelia
authors.initials: C
authors.lastname: Ferraro
authors.forename: Alba
authors.initials: A
authors.lastname: Contino
authors.forename: Andrea
authors.initials: A
authors.lastname: Scaloni
authors.forename: Stefano
authors.initials: S
authors.lastname: Donadio
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Puglia
mesh_terms: D000192:Actinomycetales / Q000235:genetics*; D022201:Chromosomes, Artificial / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D015698:Genomic Library; D013302:Streptomyces / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial