Difference between revisions of "Part:BBa K823030"
 
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<font color="#000000"; size="2">Gene clusters of ''cotZ'' and ''cgeA''</font>
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<font color="#000000"; size="2">Gene clusters ''cotVWXYZ''</font>
 
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<br>
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We compared this BioBrick with other promoters that are involved in spore crust formation such as P<sub>''cotV''</sub> and P<sub>''cgeA''</sub>.
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<br>
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We used this promoter for the expression of spore crust fusion proteins.
  
  
  
 
===Evaluation===
 
===Evaluation===
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We compared this BioBrick with other promoters that are involved in spore crust formation such as P<sub>''cotV''</sub> and P<sub>''cgeA''</sub>.
 
To test the activity we cloned this promoter into the lux-reporter vector pSBBS3C-''luxABCDE'' ([[Part:BBa_K823025|BBa_K823025]]) and measured the luminescence.
 
To test the activity we cloned this promoter into the lux-reporter vector pSBBS3C-''luxABCDE'' ([[Part:BBa_K823025|BBa_K823025]]) and measured the luminescence.
  
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http://2012.igem.org/Team:LMU-Munich/Spore_Coat_Proteins
 
http://2012.igem.org/Team:LMU-Munich/Spore_Coat_Proteins
  
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[http://2016.igem.org/Team:Freiburg Team Freiburg 2016] improved this part by including a ribosome binding site to the promoter. (see: <partinfo>BBa_K2114000</partinfo>)  <br>
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(<small>--[[User:WladStr|Wladislaw Stroukov]] 17:32, 12 October 2016 </small>)
  
  

Latest revision as of 06:49, 20 October 2016

PcotYZ: B. subtilis promoter regulating expression of spore crust proteins

The promoter PcotYZ lies within cotVWXYZ gene cluster, which regulates expression of CotYZ. The protein CotZ is a componend of the spore crust. Therefore, PcotYZ is activated in late sporulation.

Usage and Biology

Protein distribution in spore coat of Bacillus subtilis

Gene clusters cotVWXYZ



We used this promoter for the expression of spore crust fusion proteins.


Evaluation

We compared this BioBrick with other promoters that are involved in spore crust formation such as PcotV and PcgeA. To test the activity we cloned this promoter into the lux-reporter vector pSBBS3C-luxABCDE (BBa_K823025) and measured the luminescence.

The results of the luminescence measurements are shown in the following figure, which also shows the activities of the related promoters PcotV (BBa_K823033) and PcgeA.

Protein distribution in spore coat of Bacillus subtilis

The result of the measurement of this BioBrick PcotYZ and of two related promoters PcgeA and PcotV. Promoters were fused to the lux operon (pSBBS3C-luxABCDE) The left vertical axis displays the luminescence divided by the OD600 and the left one shows the OD600. On the horizontal axis the time is illustrated. The continues lines show the luminescence which correlates with the promoter activity and the dotted lines display the growth curve of each clone.


This part was used to display fusion proteins on B. subtilis spores with BBa_K823049.

Glowing spore.png


See all the details about the LMU-Munich 2012 project here: http://2012.igem.org/Team:LMU-Munich/Spore_Coat_Proteins


[http://2016.igem.org/Team:Freiburg Team Freiburg 2016] improved this part by including a ribosome binding site to the promoter. (see: BBa_K2114000)
(--Wladislaw Stroukov 17:32, 12 October 2016 )


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]