Difference between revisions of "Part:BBa K4156111"
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<partinfo>BBa_K4156111 short</partinfo>
 
<partinfo>BBa_K4156111 short</partinfo>
  
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pCaCd-mRFP is a biological brick that expresses mRFP under the control of the pCaCd promoter (BBa_K4156076). pCaCd-mRFP functions as a red fluorescent signal to characterize the performance of the pCaCd promoter.
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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.
  
  
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===Usage and Biology===
 
===Usage and Biology===
  
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In this biobrick, we used the pCaCd 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.
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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.
  
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===Characterization===
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==pH induced promoter testing==
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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.
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<html>
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<figure style="text-align:center;">
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                <img style="max-width:700px;" src="https://static.igem.wiki/teams/4156/wiki/part/1-1.png" alt="control">
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                <figcaption><b>Figure 1:</b> Induction of downstream gene mRFP expression with different pH values in different chassis organisms over 48h.</figcaption>
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              </figure>
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</html>
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<html>
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<figure style="text-align:center;">
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                <img style="max-width:700px;" src="https://static.igem.wiki/teams/4156/wiki/part/1-2-4-2.png" alt="control">
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                <figcaption><b>Figure 2:</b>  Induction of downstream gene mRFP expression over time by a pH reporter consisting of pCadC+mRFP at different pH values.</figcaption>
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              </figure>
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</html>
  
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 13:52, 10 October 2022


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.

Characterization

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.

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


control
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]