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Applied and Environmental Microbiology, June 2003, p. 3144-3151, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3144-3151.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Vector Systems Allowing Efficient Autonomous or Integrative Gene Cloning in Micromonospora sp. Strain 40027

Xiaohua Li, Xiufen Zhou, and Zixin Deng*

Bio-X Life Science Research Center, Shanghai Jiaotong University, Shanghai 200030, and School of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, People's Republic of China

Received 13 November 2002/ Accepted 25 March 2003

Vector systems allowing autonomous or site-specific integrative gene cloning were developed for Micromonospora sp. strain 40027, a producer of the antibiotic fortimicin A. The autonomous system depends on the discovery of a low-copy-number, self-transmissible covalently closed circular plasmid, pJTU112 (ca. 14.1 kb), which was shown to be present in the progenitor strain in both integrated and autonomous states. The copy numbers of both wild-type pJTU112 and three derivatives of it can be amplified at least sixfold by addition of streptomycin to the culture medium. The integrative system was developed by the use of a pBR322-derived Escherichia coli plasmid vector, pSET152, mediated by the attP site of the Streptomyces phage {Phi}C31. Both vectors can be transferred by conjugation from E. coli into Micromonospora sp. strain 40027. The heterologous cloning and expression of the dnd gene cluster originating from Streptomyces lividans 1326 into Micromonospora sp. strain 40027 demonstrated the use of the two systems.

* Corresponding author. Mailing address: Bio-X Life Science Research Center, Shanghai Jiaotong University, Shanghai 200030, China. Phone and fax: 86 21 62933404. E-mail: zxdeng{at}mail.sjtu.edu.cn.

Applied and Environmental Microbiology, June 2003, p. 3144-3151, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3144-3151.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.





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Copyright © 2003 by the American Society for Microbiology. All rights reserved.