Difference between revisions of "Part:BBa K823030"

Latest revision as of 06:49, 20 October 2016

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

The promoter P_cotYZ lies within cotVWXYZ gene cluster, which regulates expression of CotYZ. The protein CotZ is a componend of the spore crust. Therefore, P_cotYZ 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 P_cotV and P_cgeA. 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 P_cotV (BBa_K823033) and P_cgeA.

The result of the measurement of this BioBrick P_cotYZ and of two related promoters P_cgeA and P_cotV. 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.

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]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K823030 short</partinfo>
-P<sub>''cotYZ''</sub>: Promoter of ''B. subtilis'' that is involved in spore crust formation. Hence it is activated in late sporulation.
+The promoter P<sub>''cotYZ''</sub> lies within ''cotVWXYZ'' gene cluster, which regulates expression of CotYZ. The protein CotZ is a componend of the spore crust. Therefore, P<sub>''cotYZ''</sub> is activated in late sporulation.
-P<sub>''cotYZ''</sub> regulates the crust protein cotZ and cotY.
+===Usage and Biology===
 {| style="color:black;" cellpadding="3" width="60%" cellspacing="0" border="0" align="center" style="text-align:left;"
 | style="width: 60%;background-color: #EBFCE4;" |
 {|
-|[[Image:CotVWXYZ cluster.png|Protein distribution in spore coat of ''Bacillus subtilis''|400px|center]]
+|[[Image:CotVWXYZ cluster.png|Protein distribution in spore coat of ''Bacillus subtilis''|600px|center]]
 |-
 | style="width: 60%;background-color: #EBFCE4;" |
 {| style="color:black;" cellpadding="0" width="70%" cellspacing="0" border="0" align="center" style="text-align:center;"
 |style="width: 60%;background-color: #EBFCE4;" |
-<font color="#000000"; size="2">Protein distribution in spore coat of ''Bacillus subtilis''</font>
+<font color="#000000"; size="2">Gene clusters ''cotVWXYZ''</font>
 |}
 |}
 |}
+<br>
+<br>
+We used this promoter for the expression of spore crust fusion proteins.
-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.
-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>.
+===Evaluation===
+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.
+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;"
@@ Line 30: / Line 37: @@
 {| style="color:black;" cellpadding="0" width="90%" cellspacing="0" border="0" align="center" style="text-align:left;"
 |style="width: 70%;background-color: #EBFCE4;" |
-<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>
+<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>
 |}
 |}
 |}
-[[Image:Activity PcotYZ PcotV.png|thumb|center|400px|These are results of a plate reader assay to evaluate promoter activity. P<sub>''cotYZ''</sub> was cloned into [[Part:BBa_K823025|pSBBS3C-''luxABCDE'']] and luminescence was measured. This graph also shows activiy of the related promoter P''cotV'' ([[Part:BBa_K823033|BBa_K823033]]) with each two clones.]]
 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]]
@@ Line 46: / Line 52: @@
 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: <partinfo>BBa_K2114000</partinfo>)  <br>
+(<small>--[[User:WladStr|Wladislaw Stroukov]] 17:32, 12 October 2016 </small>)