Expression in Streptomyces lividans of Nonomuraea genes cloned in an artificial chromosome.
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.
Alduina
Rosa
R
Dipartimento Biologia Cellulare e dello Sviluppo, Viale delle Scienze, University of Palermo, Parco d'Orleans II, Italy. valduina@unipa.it
Giardina
Anna
A
Gallo
Giuseppe
G
Renzone
Giovanni
G
Ferraro
Clelia
C
Contino
Alba
A
Scaloni
Andrea
A
Donadio
Stefano
S
Puglia
Anna Maria
AM
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*
D004269:DNA, Bacterial