Skip to main content Accessibility help
×
Home

Construction of conjugal transfer system of Streptomyces cinnamonensis and effect of PCR-mediated nsdA gene disruption on its secondary metabolism

  • Chen Fen (a1), Xiong Wei (a2), Min Yong (a1), Fan Yu-Qing (a1), Liang Yun-Xiang (a1), Lü He-Ping (a3), Xing Ren-Chang (a3) and Zheng Ying-Hua (a1) (a3)...

Abstract

Intergeneric transfer of plasmid vectors pSET152 and pHL212 from donor Escherichia coli ET12567/pUZ8002 and S17-1 to Streptomyces cinnamonensis was demonstrated and optimized. Assisted by this conjugation system, nsdA gene disruption was achieved through PCR-targeted gene replacement. One AprRKanS exconjugant BIB309 was then isolated and confirmed to be the nsdA null mutant. Compared with the starting strain, monensin production by the nsdA mutant BIB309 increased 270% in vitro.

Copyright

Corresponding author

*Corresponding author. E-mail: zhengyh98@vip.sina.com

Footnotes

Hide All

First published in Journal of Agricultural Biotechnology 2007, 15(6): 1042–1047

Footnotes

References

Hide All
Day, LE, Chamberlin, JW, Gordee, EZ, et al. (1973) Biosynthesis of monensin. Antimicrobial Agents and Chemotherapy 4: 410414.
Gallimore, AR, Stark, CBW, Bhatt, A, et al. (2006) Evidence for the role of the monB genes, in polyether ring formation during monensin biosynthesis. Chemistry & Biology 13: 453460.
Gust, B, Challis, GL, Fowler, K, et al. (2003) PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin. Proceedings of the National Academy of Sciences, USA 100: 15411546.
Illing, GT, Normansell, DI and Peberdy, JF (1989) Protoplast isolation and regeneration in Streptomyces clavuligerus. Journal of General Microbiology 135: 22892297.
Keiser, T, Bibb, MJ, Mark, JB, et al. (2000) Practical Streptomyces Genetics. Norwich: The John Innes Foundation.
Leadlay, PF, Staunton, J, Oliynyk, M, et al. (2001) Engineering of complex polyketide biosynthesis – insights from sequencing of the monensin biosynthetic gene cluster. Journal of Industrial Microbiology & Biotechnology 27: 360367.
Li, C, Florava, G, Akopiants, K, et al. (2004) Crotonyl-coenzyme A reductase provides methylmalonyl-CoA precursors for monensin biosynthesis by Streptomyces cinnamonensis in an oil-based extended fermentation. Microbiology 150: 34633472.
Li, W, Ying, X, Guo, Y, et al. (2006) Identification of a gene negatively affecting antibiotic production and morphological differentiation in Streptomyces coelicolor A3(2). Journal of Bacteriology 188: 83688375.
Oliynyk, M, Stark, CBW, Bhatt, A, et al. (2003) Analysis of the biosynthetic gene cluster for the polyether antibiotic monensin in Streptomyces cinnamonensis and evidence for the role of monB and monC genes in oxidative cyclization. Molecular Microbiology 49: 11791190.
Sambrook, J, Fritsch, EF and Maniatis, T (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. New York: Cold Spring Harbor Laboratory Press.
Trieucuot, P, Carlier, P, Martin, P, et al. (1987) Plasmid transfer by conjugation from E. coli to gram-positive bacteria. FEMS Microbiology Letters 48: 289294.
Yu, Z, Wang, Q, Deng, Z, et al. (2006) Activation of silent antibiotic synthesis in Streptomyces lividans by disruption of a negative regulator nsdA, a gene conserved in Streptomyces. Chinese Journal of Biotechnology 22: 751762.

Keywords

Construction of conjugal transfer system of Streptomyces cinnamonensis and effect of PCR-mediated nsdA gene disruption on its secondary metabolism

  • Chen Fen (a1), Xiong Wei (a2), Min Yong (a1), Fan Yu-Qing (a1), Liang Yun-Xiang (a1), Lü He-Ping (a3), Xing Ren-Chang (a3) and Zheng Ying-Hua (a1) (a3)...

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed.