Difference between revisions of "Part:BBa K823030"
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We used the promtoer P<sub>''cotYZ''</sub>, P<sub>''cotV''</sub> and P<sub>''cgeA''</sub> for the regulation of spore crust proteins in ''B. subtilis''.  
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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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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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These are results of a plate reader assay to evaluate promoter activity. Promoters were cloned into [[Part:BBa_K823025|pSBBS3C-''luxABCDE'']] and luminescence was measured. This graph also shows activiy of the related promoter P<sub>''cotV''</sub> ([[Part:BBa_K823033|BBa_K823033]]) and P<sub>''cgeA''</sub>.
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The results of the luminescence measurements are shown in the following figure, which also shows the activities of the related promoters P<sub>''cotV''</sub> ([[Part:BBa_K823033|BBa_K823033]]) and P<sub>''cgeA''</sub>.
  
 
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<font color="#000000"; size="2"><p align="justify">The result of the measurement of the three promoters P<sub>''cgeA''</sub>, P<sub>''cotV''</sub> and P<sub>''cotYZ''</sub>. The left vertical axis displays the luminescence divided by the OD<sub>600</sub> and the left one shows the OD<sub>600</sub>. On the horizontal axis the time is illustrated. The drawn through lines show the luminescence which correlates with the promoter activity and the dotted lines display the growth curve of each clone.</p></font>
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<font color="#000000"; size="2"><p align="justify">The result of the measurement of this BioBrick P<sub>''cotYZ''</sub> and of two related promoters P<sub>''cgeA''</sub> and P<sub>''cotV''</sub>. Promoters were fused to the lux operon ([[Part:BBa_K823025|pSBBS3C-''luxABCDE'']]) The left vertical axis displays the luminescence divided by the OD<sub>600</sub> and the left one shows the OD<sub>600</sub>. 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.</p></font>
 
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Revision as of 21:54, 26 September 2012

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

PcotYZ: Promoter of B. subtilis that is involved in spore crust formation. Hence it is activated in late sporulation. PcotYZ regulates the crust protein cotZ and cotY.

Usage and Biology

Protein distribution in spore coat of Bacillus subtilis

Gene clusters of cotZ and cgeA

We compared this BioBrick with other promoters that are involved in spore crust formation such as PcotV and PcgeA.


Evaluation

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



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]