Difference between revisions of "Part:BBa K823033"
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<partinfo>BBa_K823033 short</partinfo>
 
<partinfo>BBa_K823033 short</partinfo>
  
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P<sub>''cotV''</sub>: Promoter of ''B. subtilis'' that is involved in spore crust formation. Hence it is activated in late sporulation.
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The promoter P<sub>''cotV''</sub> lies within ''cotVWXYZ'' gene cluster, which regulates expression of CotV. The protein CotV is a componend of the spore crust. Therefore, P<sub>''cotV''</sub> is activated in late sporulation.
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P<sub>''cotV''</sub> regulates the crust protein ''cotV''.
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===Usage and Biology===
 
===Usage and Biology===
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{| style="color:black;" cellpadding="0" width="70%" cellspacing="0" border="0" align="center" style="text-align:center;"
 
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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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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>''cotYZ''</sub> and P<sub>''cgeA''</sub>.
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===Evaluation===
 
===Evaluation===
 
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>''cotYZ''</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>.
  
 
{| style="color:black;" cellpadding="3" width="70%" cellspacing="0" border="0" align="center" style="text-align:left;"
 
{| style="color:black;" cellpadding="3" width="70%" cellspacing="0" border="0" align="center" style="text-align:left;"
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{| style="color:black;" cellpadding="0" width="90%" cellspacing="0" border="0" align="center" style="text-align:left;"
 
{| style="color:black;" cellpadding="0" width="90%" cellspacing="0" border="0" align="center" style="text-align:left;"
 
|style="width: 70%;background-color: #EBFCE4;" |
 
|style="width: 70%;background-color: #EBFCE4;" |
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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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This part was used to display fusion proteins on ''B. subtilis'' spores. However we experienced better results by using P<sub>''cotYZ''</sub> ([[Part:BBa K823030|Part:BBa K823030]], see device [[Part:BBa_K823049|BBa_K823049]]).
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This part was used to display fusion proteins on ''B. subtilis'' spores with [[Part:BBa_K823049|BBa_K823049]].
  
 
[[Image:Glowing spore.png|100px|link=Part:BBa_K823049]]
 
[[Image:Glowing spore.png|100px|link=Part:BBa_K823049]]
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See all the details about the LMU-Munich 2012 project here:
 
See all the details about the LMU-Munich 2012 project here:
 
http://2012.igem.org/Team:LMU-Munich/Spore_Coat_Proteins
 
http://2012.igem.org/Team:LMU-Munich/Spore_Coat_Proteins
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 22:42, 26 September 2012

PcotV: B. subtilis promoter regulates spore crust protein cotV

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

Usage and Biology

Protein distribution in spore coat of Bacillus subtilis

Gene clusters cotVWXYZ.

We compared this BioBrick with other promoters that are involved in spore crust formation such as PcotYZ 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]