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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: zhengyh98@vip.sina.com
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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

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Footnotes

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

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