대한의생명과학회지 (Biomedical Science Letters)

  • 제18권1호
  • /
  • Pages.29-34
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  • 2012
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  • 1738-3226(pISSN)
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  • 2288-7415(eISSN)
대한의생명과학회 (The Korean Society for Biomedical Laboratory Sciences)
권호 표지

Construction of Expression Vector for Functional Analysis of Target Protein in Streptomyces sp.

  • Lee, Yong-Jik (System & Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Ryu, Jae-Ki (Department of Biomedical Laboratory Science, Gimcheon University) ;
  • Kim, Hyun-Soo (Department of Microbiology, Keimyung University)
  • 투고 : 2011.10.06
  • 심사 : 2012.03.25
  • 발행 : 2012.03.31
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초록

Streptomycetes are gram-positive filamentous bacteria that are well-known for producing a vast array of bioactive compounds, including more than 70 % of commercially important antibiotics. For the research about Streptomyces sp., the protoplast and electroporation transformation method have been the general techniques for the construction of transformants. However, these techniques have low efficiency and are time-consuming. Another option is intergenic conjugation, which is used for DNA transfer using methylation-deficient E. coli as a DNA donor to avoid the methylated-DNA-dependent restriction systems of actinomycetes. This conjugation method has been widely improved and applied to many other actinomycetes. In this research, an effective transformation procedure for the construction of expression vector by using gateway system was established to avoid limit of restriction enzyme site for cloning of target gene based on transconjugation by Escherichia coli ET12567/pUZ8002 with a pSET152 integration vector.

키워드

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