Hostname: page-component-78c5997874-g7gxr Total loading time: 0 Render date: 2024-11-12T19:44:21.050Z Has data issue: false hasContentIssue false
  • English
  • Français

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

Published online by Cambridge University Press:  27 June 2008

Chen Fen ,
Xiong Wei ,
Min Yong ,
Fan Yu-Qing ,
Liang Yun-Xiang ,
Lü He-Ping ,
Xing Ren-Chang  and
Zheng Ying-Hua
Chen Fen
Affiliation:
National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China
Xiong Wei
Affiliation:
Department of Biological Science and Biotechnology, Tsinghua University, Beijing 100084, China
Min Yong
Affiliation:
National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China
Fan Yu-Qing
Affiliation:
National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China
Liang Yun-Xiang
Affiliation:
National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China
Lü He-Ping
Affiliation:
Beijing Institute of Biomedicine, Beijing 100091, China
Xing Ren-Chang
Affiliation:
Beijing Institute of Biomedicine, Beijing 100091, China
Zheng Ying-Hua*
Affiliation:
National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China Beijing Institute of Biomedicine, Beijing 100091, China
*
*Corresponding author. E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

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.

Keywords

conjugal transfernsdAPCR-mediated gene disruptionStreptomyces cinnamonensis
Type
Research Papers
Information
Chinese Journal of Agricultural Biotechnology , Volume 5 , Issue 1 , April 2008 , pp. 73 - 79
Copyright
Copyright © China Agricultural University 2008

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

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

References

Day, LE, Chamberlin, JW, Gordee, EZ, et al. (1973) Biosynthesis of monensin. Antimicrobial Agents and Chemotherapy 4: 410–414.CrossRefGoogle ScholarPubMed
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: 453–460.CrossRefGoogle ScholarPubMed
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: 1541–1546.CrossRefGoogle ScholarPubMed
Illing, GT, Normansell, DI and Peberdy, JF (1989) Protoplast isolation and regeneration in Streptomyces clavuligerus. Journal of General Microbiology 135: 2289–2297.Google ScholarPubMed
Keiser, T, Bibb, MJ, Mark, JB, et al. (2000) Practical Streptomyces Genetics. Norwich: The John Innes Foundation.Google Scholar
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: 360–367.CrossRefGoogle ScholarPubMed
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: 3463–3472.CrossRefGoogle Scholar
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: 8368–8375.CrossRefGoogle ScholarPubMed
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: 1179–1190.CrossRefGoogle ScholarPubMed
Sambrook, J, Fritsch, EF and Maniatis, T (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. New York: Cold Spring Harbor Laboratory Press.Google Scholar
Trieucuot, P, Carlier, P, Martin, P, et al. (1987) Plasmid transfer by conjugation from E. coli to gram-positive bacteria. FEMS Microbiology Letters 48: 289–294.CrossRefGoogle Scholar
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: 751–762.Google ScholarPubMed