Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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P22-Based Challenge Phage Constructs to Study Protein-Protein Interactions between the $\sigma$$^{54}$-Dependent Promoter, dctA, and Its Transcriptional Regulators

  • Song, Jeong-Min (Department of Ophthalmology and The Institute of Vision Research, Yonsei University College of Medicine) ;
  • Kim, Eungbin (Department of Biology and Institute of Bioscience and Biotechnology, Yonsei University) ;
  • Lee, Joon H. (Protein Netwark Research Center, Yonsei University)
  • Published : 2002.09.01
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Abstract

To study interactions between $C_{4}$-dicarboxylic acid transport protein D and E$\sigma$$^{54}$ in the dctA promoter regulatory region, we used the challenge phage system. An ant＇-｀lac fusion was recombined onto the challenge phage, and this ant＇-｀lac fusion along with Pant and the R. meliloti dctA promoter regulatory region were cloned onto a plasmid. The plasmid bearing the ant＇-｀lac fusion was used as a reporter plasmid in a coupled transcription-translation system. Addition of purified $\sigma$$^{54}$ to the coupled system specifically repressed transcription of the plasmid-borne ant＇-｀lac fusion. When DCTD was added along with $\sigma$$^{54}$ to the coupled system, transcription of the ant＇-｀lac fusion was even further repressed, suggesting that DCTD may stabilize closed complexes between E$\sigma$$^{54}$ and the dctA promoter.

Keywords

References

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