Part:BBa_K2317000

wt-Pr

Wt-Pr is a E.coil constitutive weak promoter isolated from a coding part BBa_K1031211 submitted by Peking 2013 iGEM.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Description

Wt-Pr is the natural promoter upstream DmpR from dmp operon in Pseudomonas sp. Strain CF600 [1,2]. Reported activity of the promoter is given as the EGFP of the PET28a plasmid in strain BL21 grown in LB media.

To figure out whether EGFP expression will have effects on bacteria growth, we tested the growth rate of different constructed bacteria containing different promoters. The growing tendency of bacteria expressing EGFP was similar with unloaded vector within 12 hours, which suggested that in 12 hours, the expression of EGFP wouldn’t have effects on the growing of bacteria (Fig 1A).

We used the ratio of EGFP and value of OD600 to make comparisons on promoter strength. It turned out that promotor J23101 expressed strongest among all the promotors, and J23107 next. Pr promotor expressed week eGFP, and showed no difference with J23114 (Part J23114 is a member of Anderson family of constitutive promoter. It is a much weaker member in this family )in expression level (Fig 1B).

Figure 1. Promoter strength certification. (A) Growth curve of different constructed bacteria. Constructions contained promoter J23101, J23107, J23114, wt-Pr and mock (B) ratio of EGFP/ OD600

Reference

[1] Shingler, V., M. Bartilson, and T. Moore. Cloning and nucleotide sequence of the gene encoding the positive regulator (DmpR) of the phenol catabolic pathway encoded by pVI150 and identification of DmpR as a member of the NtrC family of transcriptional activators. J. Bacteriol. 1993,175: 1596–1604.

[2] Shingler, V., and T. Moore. Sensing of aromatic compounds by the DmpR transcriptional activator of phenol-catabolizing Pseudomonas sp. strain CF600. J. Bacteriol. 1994, 176:1555–1560.