Difference between revisions of "Part:BBa K823030"

Revision as of 16:20, 26 September 2012

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

P_cotYZ: Promoter of B. subtilis that is involved in spore crust formation. Hence it is activated in late sporulation. P_cotYZ 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 used the promtoer P_cotYZ, P_cotV and P_cgeA for the regulation of spore crust proteins in B. subtilis.

Evaluation

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 P_cotV (BBa_K823033) and P_cgeA.

The result of the measurement of the three promoters P_cgeA, P_cotV and P_cotYZ. 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 with BBa_K823049.

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]

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 P<sub>''cotYZ''</sub> regulates the crust protein cotZ and cotY.
+===Usage and Biology===
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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.
+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''.
+===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.
 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>.