Difference between revisions of "Part:BBa K823003"

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===Usage and Biology===
 
===Usage and Biology===
 
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<p align="justify">P<sub>''veg''</sub> 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<sub>''veg''</sub> 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 . </p>
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<p align="justify">P<sub>''veg''</sub> 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<sub>''veg''</sub> 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 '''Bead'''zillus]. </p>
 
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Revision as of 08:22, 26 September 2012

Pveg

Pveg is a strong, constitutive promoter of Bacillus subtilis.


Usage and Biology


Pveg 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)]. Pveg 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

β-galactosidase assay


The two constitutive promoters PliaG and Pveg were evaluated with the reporter vector pSBBs1C-lacZ which contains the lacZ reporter gene (Fig.1).


Englisch Auswertung PliaG Pveg.png

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 Pveg and PliaG 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 Pveg and PliaG is very similar based on the growth curve. The highest β-galactosidase activity and therefore the highest activity of the promoter Pveg 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 PliaG 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]