Difference between revisions of "Part:BBa K4156110"
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==hypoxia induced promoter testing==
 
==hypoxia induced promoter testing==
  
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We constructed a hypoxia reporter consisting of the hypoxia-inducible promoter pPepT+mRFP. To test its performance, we added reporter in different chassis organisms.Fig 1,2 indicates that pPepT induces the expression of the downstream gene mRFP with the decrease of O2. Thus, it can be seen that the hypoxia reporter can work properly.
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We constructed a hypoxia reporter consisting of the hypoxia-inducible promoter pPepT+mRFP. To test its performance, we added reporter in different chassis organisms.Fig 1 indicates that pPepT induces the expression of the downstream gene mRFP with the decrease of O2. Thus, it can be seen that the hypoxia reporter can work properly.
  
 
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                <figcaption><b>Figure 2:</b> Induction of downstream gene mRFP expression over time by a hypoxia reporter consisting of pPepT+mRFP under hypoxic and normoxic conditions.</figcaption>
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 13:46, 11 October 2022


pPepT-mRFP

pPepT-mRFP is a biological brick that expresses mRFP controlled by the pPepT promoter (BBa_K4156110). pPepT-mRFP functions to characterize the performance of the pPepT promoter using a red fluorescent signal.


Usage and Biology

In this biobrick, we used the pPepT promoter to regulate the expression of downstream mRFP 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 pPepT promoter to be measured by the red fluorescent signal. We were thus able to verify the specific response of the pPepT promoter to changes in oxygen concentration.

Characterization

hypoxia induced promoter testing

We constructed a hypoxia reporter consisting of the hypoxia-inducible promoter pPepT+mRFP. To test its performance, we added reporter in different chassis organisms.Fig 1 indicates that pPepT induces the expression of the downstream gene mRFP with the decrease of O2. Thus, it can be seen that the hypoxia reporter can work properly.

control
Figure 1: Induction of downstream gene mRFP expression under hypoxic and normoxic conditions in different chassis organisms over 48h.


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 723
    Illegal AgeI site found at 835
  • 1000
    COMPATIBLE WITH RFC[1000]