Part:BBa_K823030

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 component of the spore crust. Therefore, P_cotYZ is activated in late sporulation.

Usage and Biology

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 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.

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]