| Title | P22-Based Challenge Phage Constructs to Study Protein-Protein Interactions between the [sigma]^54 -Dependent Promoter, dctA, and Its Transcriptional Regulators |
|---|---|
| Author | Jeong Min Song 1 , Eungbin Kim 2 , and Joon H. Lee 1,3 * |
| Address | 1 Department of Ophthalmology and The Institute of Vision Research, Yonsei University, College of Medicine, Seoul 120-749, Korea; 2 Department of Biol |
| Bibliography | Journal of Microbiology, 40(3),205-210, 2002, |
| DOI | |
| Key Words | challenge phage system, [sigma]^54 , dctA, DCTD |
| 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. |
| Download PDF | 403-03.pdf |