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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PcotV: B. subtilis promoter regulates spore crust proteins and is activated in late sporulation. PcotYZ regulates the crust protein cotZ and cotY. We used this promoter to control the fusion proteins for display on B. subtilis spores.
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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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P<sub>''cotV''</sub> regulates the crust protein ''cotV''.
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{| style="color:black;" cellpadding="3" width="60%" cellspacing="0" border="0" align="center" style="text-align:left;"
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| style="width: 60%;background-color: #EBFCE4;" |
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{|
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|[[Image:CotVWXYZ cluster.png|Protein distribution in spore coat of ''Bacillus subtilis''|400px|center]]
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<font color="#000000"; size="2">Gene clusters of ''cotZ'' and ''cgeA''</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''. 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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{| style="color:black;" cellpadding="3" width="70%" cellspacing="0" border="0" align="center" style="text-align:left;"
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| style="width: 70%;background-color: #EBFCE4;" |
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{|
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|[[Image:Activity PcotYZ PcotV.png|Protein distribution in spore coat of ''Bacillus subtilis''|610px|center]]
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{| style="color:black;" cellpadding="0" width="90%" cellspacing="0" border="0" align="center" style="text-align:left;"
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|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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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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[[Image:Glowing spore.png|100px|link=Part:BBa_K823049]]
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We experienced weaker strength as the related promoter PcotYZ [http://2012.igem.org/Team:LMU-Munich/Spore_Coat_Proteins Part:BBa_K823030]
 
  
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see all the details about the LMU-Munich 2012 project here: http://2012.igem.org/Team:LMU-Munich/Spore_Coat_Proteins
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See all the details about the LMU-Munich 2012 project here:
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http://2012.igem.org/Team:LMU-Munich/Spore_Coat_Proteins
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 15:27, 26 September 2012

PcotV: B. subtilis promoter regulates spore crust protein cotV

PcotV: Promoter of B. subtilis that is involved in spore crust formation. Hence it is activated in late sporulation. PcotV regulates the crust protein cotV.

Protein distribution in spore coat of Bacillus subtilis

Gene clusters of cotZ and cgeA

We used the promtoer PcotYZ, PcotV and PcgeA for the regulation of spore crust proteins in B. subtilis. To test the activity we cloned this promoter into the lux-reporter vector pSBBS3C-luxABCDE (BBa_K823025) and measured the luminescence.

These are results of a plate reader assay to evaluate promoter activity. Promoters were cloned into pSBBS3C-luxABCDE and luminescence was measured. This graph also shows activiy of the related promoter PcotV (BBa_K823033) and PcgeA.

Protein distribution in spore coat of Bacillus subtilis

The result of the measurement of the three promoters PcgeA, PcotV and PcotYZ. 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 drawn through 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. However we experienced better results by using PcotYZ (Part:BBa K823030, see device 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]