Engineering Streptomyces coelicolor for heterologous expression of secondary metabolite gene clusters
Version of Record online: 26 OCT 2010
© 2010 The Authors. Journal compilation © 2010 Society for Applied Microbiology and Blackwell Publishing Ltd
Special Issue: Crystal ball and Streptomyces Special issue. Guest Editors: Hildgund Schrempf, Paul Dyson and Sergey Zotchev
Volume 4, Issue 2, pages 207–215, March 2011
How to Cite
Gomez-Escribano, J. P. and Bibb, M. J. (2011), Engineering Streptomyces coelicolor for heterologous expression of secondary metabolite gene clusters. Microbial Biotechnology, 4: 207–215. doi: 10.1111/j.1751-7915.2010.00219.x
- Issue online: 22 FEB 2011
- Version of Record online: 26 OCT 2010
- Received 10 July, 2010; accepted 3 September, 2010.
Fig. S1. Genomotyping analysis. (A) Scatter plot of normalized data from the quadruple deletion mutant (QDM) M1146 (Y-axis) versus M145 (X-axis). (B) Scatter plot of normalized data from M1154 (Y-axis) versus M145 (X-axis). In green, positive controls (fragments used for homologous hybridization); in red, negative controls (deleted genes); in grey, rest of the genes; diagonal lines from top to bottom, two-fold increase in intensity, 1:1 ratio and two-fold decrease in intensity. (C) Southern blot of digested DNA from M145 and M1146 hybridized with a probe derived from some of the genes deleted from the cda gene cluster that gave higher than expected intensities upon genomotyping. The expected fragments are detected only in M145 and not in M1146, while several cross-hybridizing fragments are observed in both strains.
Fig. S2. Growth and differentiation. (A) Growth and sporulation on SFM agar medium. Plates were scanned from above (top row) and below (bottom row; the image was inverted horizontally for easier interpretation) at the indicated times. Note the delayed growth and reduction in spore pigmentation in M1152 and M1154. (B) Antimicrobial activity detected in M145 but not in M1154 GYM culture supernatants.
Fig. S3. Identification of congocidine. MS/MS analysis of a fraction of the large peak eluting at 10.5 min in the chromatogram from M1154 containing the congocidine gene cluster (Fig. 4). 431 m z−1 corresponds to congocidine + 1 hydrogen ion ; 453 m z−1 corresponds to the sodium adduct; 216 m z−1 corresponds to doubly charged congocidine + 2 hydrogen ions; See Juguet and colleagues (2009) for further explanation of the MS/MS data and fractionation spectra.
Table S1. Oligonucleotides used in this study.
Table S2. Vectors and constructs used in this study.
Table S3. Strains used in this study.
Table S4. Data for Fig. 3 for chloramphenicol (CP) (Fig. 3A) and congocidine (CGC) (Fig. 3B) production. Strains were grown in liquid GYM medium inoculated with pre-germinated spores (chloramphenicol gene cluster) or with mycelium from a seed culture (congocidine gene cluster).
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