Expression in Streptomyces lividans of Nonomuraea genes cloned in an artificial chromosome
| Autor: | Anna Giardina, Andrea Scaloni, Anna Maria Puglia, Giuseppe Gallo, Rosa Alduina, Alba Contino, Giovanni Renzone, Stefano Donadio, Clelia Ferraro |
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| Přispěvatelé: | Alduina R, Giardina A, Gallo G, Renzone G, Ferraro C, Contino A, Scaloni A, Donadio S, Puglia AM |
| Rok vydání: | 2005 |
| Předmět: |
DNA
Bacterial Chromosomal library of Nonomuraea sp. ATCC39727 Escherichia coli–Streptomyces artificial chromosome (ESAC) RT-PCR Molecular cloning Applied Microbiology and Biotechnology Streptomyces Genetic analysis Thiostrepton chemistry.chemical_compound Actinomycetales Chromosomes Artificial Cloning Molecular A40926 Gene Regulator gene Genetics Genomic Library biology MALDI-TOF mass spectrometry Promoter General Medicine biology.organism_classification dbv gene cluster 2D-PAGE chemistry Genes Bacterial Heterologous expression Pulsed field gel electrophoresi dalbavancin Biotechnology |
| Zdroj: | Applied Microbiology and Biotechnology. 68:656-662 |
| ISSN: | 1432-0614 0175-7598 |
| DOI: | 10.1007/s00253-005-1929-y |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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