Part:BBa_K823003

P_veg

P_veg is a strong, constitutive promoter of Bacillus subtilis.

Usage and Biology

P_veg is known to show a strong constitutive activity during the vegetative growth phase and sporulation. This promoter is important for the transcription of the veg gene, which plays a role during sporulation [http://www.ncbi.nlm.nih.gov/pubmed?term=J.%20Biochem.%2C%20133%20%284%29%3A%20475%E2%80%93483: (Fukushima et al., 2003)]. P_veg was measured by using the reporter gene lacZ. For more Details visit the [http://2012.igem.org/Team:LMU-Munich/Data/Constitutive Data of the constitutive promoters] of the LMU-Munich Team 2012 or get an overview of our whole project [http://2012.igem.org/Team:LMU-Munich Beadzillus].

Evaluation

This part was also evaluated in the publication [http://www.jbioleng.org/content/7/1/29 The Bacillus BioBrick Box: generation and evaluation of essential genetic building blocks for standardized work with Bacillus subtilis] by Radeck et al..

β-galactosidase assay

The two constitutive promoters P_liaG and P_veg were evaluated with the reporter vector pSB_Bs1C-lacZ which contains the lacZ reporter gene (Fig.1).

Fig. 1: β-galactosidase assay and growth curve of strains carrying the promoters PliaG (black) and Pveg (grey) fused to lacZ. β-galactosidase activity (Miller Units) and the growth curve values are the average of two independent clones with their standard deviation. Experiment shows representative data from three independent experiments.

Promoter activity leads to the expression of the β-galactosidase. The β-galactosidase assay of the constitutive Bacillus promoters P_veg and P_liaG was repeated three times. The graph shows data of one representative experiment. In the beginning of the growth curve both promoters show only low activity before it increases to a maximum before it decreases to the begininng level after about seven hours (Data not shown). Summing up, the course of activity of both promoters P_veg and P_liaG is very similar based on the growth curve. The highest β-galactosidase activity and therefore the highest activity of the promoter P_veg with a maximum of 65 Miller units can be found during the transition from the logarithmic to the stationary phase. This is about five times higher than the acitivity of the promoter P_liaG with a maximum activity of about 12 Miller Units.

Sequence and Features

This part was amplified from the genom of B. subtilis.

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]