Essential role of an unknown gene aziU3 in the production of antitumor antibiotic azinomycin B verified by utilizing optimized genetic manipulation systems for Streptomyces sahachiroi
Article first published online: 2 NOV 2012
© 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved
FEMS Microbiology Letters
Volume 337, Issue 2, pages 147–154, December 2012
How to Cite
Wang, S., Zhao, R., Liu, K., Zhu, M., Li, A. and He, J. (2012), Essential role of an unknown gene aziU3 in the production of antitumor antibiotic azinomycin B verified by utilizing optimized genetic manipulation systems for Streptomyces sahachiroi. FEMS Microbiology Letters, 337: 147–154. doi: 10.1111/1574-6968.12020
- Issue published online: 28 NOV 2012
- Article first published online: 2 NOV 2012
- Accepted manuscript online: 8 OCT 2012 05:01AM EST
- Manuscript Accepted: 2 OCT 2012
- Manuscript Revised: 21 SEP 2012
- Manuscript Received: 25 JUL 2012
- Natural Science Foundation of China. Grant Numbers: 30800020, 30970059
- Ministry of Education of China. Grant Number: NECT-08-0779
- Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry. Grant Number: 1590
- Fundamental Research Funds for the Central Universities. Grant Numbers: 2009PY006, CCNU10A02011
Data S1. Materials and methods.
Fig. S1. Effect of various liquid media on S. sahachiroi protoplast formation and regeneration.
Fig. S2. Effect of culture time on S. sahachiroi protoplast formation and regeneration.
Fig. S3. Effects of lysozyme concentration and reaction time on S. sahachiroi protoplast formation and regeneration.
Fig. S4. Effect of recipient/donor ratio on conjugation efficiency.
Fig. S5. Effect of media on sporulation of S. sahachiroi.
Fig. S6. Conjugation and transformation plates.
Fig. S7. PCR confirmation of the in-frame deletion mutant ΔaziU3 and the complementation mutant ΔaziU3::aziU3.
Fig. S8. PFGE analysis of the native linear plasmids in S. sahachiroi ATCC 33158.
Table S1. Bacterial strains and plasmids used in this study.
Table S2. PCR primers used in this study.
Table S3. Sensitivity of S. sahachiroi ATCC 33158 to antibiotics.
Table S4. Effect of culture temperature on protoplast formation and regeneration.
Table S5. Effect of regeneration media on protoplast regeneration.
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