Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Two pHZ1358 Derivative Vectors for Efficient Gene Knockout in Streptomyces

  • He, Yunlong (Laboratory of Microbial Metabolism, and School of Life Sciences and Biotechnology, Shanghai Jiaotong University) ;
  • Wang, Zhijun (Laboratory of Microbial Metabolism, and School of Life Sciences and Biotechnology, Shanghai Jiaotong University) ;
  • Bai, Linquan (Laboratory of Microbial Metabolism, and School of Life Sciences and Biotechnology, Shanghai Jiaotong University) ;
  • Liang, Jingdan (Laboratory of Microbial Metabolism, and School of Life Sciences and Biotechnology, Shanghai Jiaotong University) ;
  • Zhou, Xiufen (Laboratory of Microbial Metabolism, and School of Life Sciences and Biotechnology, Shanghai Jiaotong University) ;
  • Deng, Zixin (Laboratory of Microbial Metabolism, and School of Life Sciences and Biotechnology, Shanghai Jiaotong University)
  • Received : 2009.10.22
  • Accepted : 2009.11.28
  • Published : 2010.04.28
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Abstract

The deletion of sti from the Streptomyces plasmid pIJ101 made its derivative pHZ1358 an efficient vector for gene disruption and replacement. Here, pHZ1358 was further optimized by the construction of a derivative plasmid pJTU1278, in which a cassette carrying multiple cloning sites and a lacZ selection marker were introduced for convenient plasmid construction in E. coli. In addition, the oriT region of pJTU1278 was also deleted, generating a vector (pJTU1289) that can be used specifically for PCR-targeting. The efficient usage of these vectors was demonstrated by the deletion of the gene involved in avermectin biosynthesis in S. avermitilis.

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References

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