TY - JOUR
T1 - P22-based challenge phage constructs to study protein-protein interactions between the σ54-dependent promoter, dctA, and its transcriptional regulators
AU - Song, Jeong Min
AU - Kim, Eungbin
AU - Lee, Joon H.
N1 - Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2002/9
Y1 - 2002/9
N2 - To study interactions between C4-dicarboxylic acid transport protein D and Eσ54 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 σ54 to the coupled system specifically repressed transcription of the plasmid-borne ant'-'lac fusion. When DCTD was added along with σ54 to the coupled system, transcription of the ant'-'lac fusion was even further repressed, suggesting that DCTD may stabilize closed complexes between Eσ54 and the dctA promoter.
AB - To study interactions between C4-dicarboxylic acid transport protein D and Eσ54 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 σ54 to the coupled system specifically repressed transcription of the plasmid-borne ant'-'lac fusion. When DCTD was added along with σ54 to the coupled system, transcription of the ant'-'lac fusion was even further repressed, suggesting that DCTD may stabilize closed complexes between Eσ54 and the dctA promoter.
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M3 - Article
AN - SCOPUS:0036750938
VL - 40
SP - 205
EP - 210
JO - Journal of Microbiology
JF - Journal of Microbiology
SN - 1225-8873
IS - 3
ER -