Part:BBa_K4156111

pCadC-mRFP

pCadC-mRFP is a biological brick that expresses mRFP under the control of the pCaCd promoter (BBa_K4156076). pCadC-mRFP functions as a red fluorescent signal to characterize the performance of the pCaCd promoter.

Usage and Biology

In this biobrick, we used the pCadC promoter to regulate downstream mRFP expression and eventually introduced the rrnB T1 and T7Te dual terminators. in addition, we added the RiboJ sequence and RBS B0034 for optimization. This design allowed the spatiotemporal intensity of gene expression under the regulation of the pCaCd promoter to be measured by the red fluorescent signal. We were thus able to verify the specific response of the pCaCd promoter to pH changes.

pH induced promoter testing

We constructed a pH reporter consisting of the pH-inducible promoter pCadC+mRFP. To test itsperformance, we added reporter in different chassis organisms. Fig 1,2 illustrates that pCadC induces the expression of the downstream gene mRFP with the decrease of pH,. Thus, it can be seen that pH reporter can work properly.

Figure 1: Induction of downstream gene mRFP expression with different pH values in different chassis organisms over 48h.

Figure 2: Induction of downstream gene mRFP expression over time by a pH reporter consisting of pCadC+mRFP at different pH values.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 731
  Illegal AgeI site found at 843
  
• 1000
  COMPATIBLE WITH RFC[1000]