Difference between revisions of "Part:BBa K2832180"

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== Measurements ==
 
== Measurements ==
  
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BBa_K2832180 (apFAB51) was used in the [https://parts.igem.org/Part:BBa_K3792005 apFAB51 Measurement Device (BBa_K3792005)]. The biological device was inserted in the pSB1K3 plasmid backbone resulting in pFSUIGEM4. Figure 1 is the map of pFSUIGEM4. E. coli NEBExpress was transformed with pFSUIGEM4 resulting in the apFAB51 Measurement Cell.
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BBa_K2832180 (apFAB51) was used in the [https://parts.igem.org/Part:BBa_K3792005 apFAB51 Measurement Device (BBa_K3792005)]. The biological device was inserted in the pSB1K3 plasmid backbone resulting in pFSUIGEM4. Figure 1 depicts the map of pFSUIGEM4.  
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apFAB51 Measurement Cell was grown in LB Broth medium then fluorescence was measured in a flow cytometer. The results of the measurements are depicted in Figure 2.
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   <figcaption>Figure 1 - pFSUIGEM4 Map.</figcaption>
 
   <figcaption>Figure 1 - pFSUIGEM4 Map.</figcaption>
 
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E. coli NEBExpress was transformed with pFSUIGEM4 resulting in the apFAB51 Measurement Cell. apFAB51 Measurement Cell was grown in LB Broth medium then fluorescence was measured in a flow cytometer. The results of the measurements are depicted in Figure 2.
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     <img src="https://static.igem.wiki/teams/4680/wiki/contributions/negative-control-cell-vs-apfab51-test-cell-1.jpeg" alt="apFAB51 Measurement Cell Flow Cytometry Results" width="100%">
 
     <img src="https://static.igem.wiki/teams/4680/wiki/contributions/negative-control-cell-vs-apfab51-test-cell-1.jpeg" alt="apFAB51 Measurement Cell Flow Cytometry Results" width="100%">
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    <figcaption>Figure 2 - apFAB51 Measurement Cell Flow Cytometry Results.</figcaption>
 
 
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  <figcaption>Figure 2 - apFAB51 Measurement Cell Flow Cytometry Results.</figcaption>
 
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In Figure 2, the upper row is the results of a Negative Control Cell that is the same chassis as the apFAB51 Measurement Cell (E. coli NEBExpress). The plasmid vector is pSB1K3 with an inserted sequence of DNA that has no function. The lower row is the results of the apFAB51 Measurement Cell. The first column of graphs (A1 and A2) are the scatterplots of the flow cytometer events and the subpopulation of events that were selected for further analysis. The second column (B1 and B2) are the scatterplots of the fluorescent emission after excitation of the cells. The third column (C1 and C2) are histograms with fluorescence in the horizontal axis and frequency in the vertical axis. The results indicate that the apFAB51 Measurement Cell is slightly more fluorescent than the Negative Control Cell. The results are not consistent with what we can observe with the naked eye where the apFAB51 Measurement Cells look red in ambient light while the Negative Control Cells do not. We are repeating these measurements and using the iGEM recommended protocol for calibrating the fluorescence. We will present the results of the second set of measurements at the iGEM Grand Jamboree.
  
  

Latest revision as of 18:33, 11 October 2023


BIOFAB Modular Promoter apFAB51

BBa_K2832180 is apFAB51 in the BIOFAB Collection . The BIOFAB Collection has constitutive promoters and transcription terminators. The collection is curated by the FSU iGEM teams . BBa_K2832180 (apFAB51) is an engineered constitutive promoter. Information about the design of this promoter is available in the BIOFAB Collectionpage. The FSU iGEM teams are characterizing all the parts in the BIOFAB Collection so they are useful to the iGEM community

Measurements

BBa_K2832180 (apFAB51) was used in the apFAB51 Measurement Device (BBa_K3792005). The biological device was inserted in the pSB1K3 plasmid backbone resulting in pFSUIGEM4. Figure 1 depicts the map of pFSUIGEM4.

pFSUIGEM4 Map
Figure 1 - pFSUIGEM4 Map.

E. coli NEBExpress was transformed with pFSUIGEM4 resulting in the apFAB51 Measurement Cell. apFAB51 Measurement Cell was grown in LB Broth medium then fluorescence was measured in a flow cytometer. The results of the measurements are depicted in Figure 2.

apFAB51 Measurement Cell Flow Cytometry Results
Figure 2 - apFAB51 Measurement Cell Flow Cytometry Results.

In Figure 2, the upper row is the results of a Negative Control Cell that is the same chassis as the apFAB51 Measurement Cell (E. coli NEBExpress). The plasmid vector is pSB1K3 with an inserted sequence of DNA that has no function. The lower row is the results of the apFAB51 Measurement Cell. The first column of graphs (A1 and A2) are the scatterplots of the flow cytometer events and the subpopulation of events that were selected for further analysis. The second column (B1 and B2) are the scatterplots of the fluorescent emission after excitation of the cells. The third column (C1 and C2) are histograms with fluorescence in the horizontal axis and frequency in the vertical axis. The results indicate that the apFAB51 Measurement Cell is slightly more fluorescent than the Negative Control Cell. The results are not consistent with what we can observe with the naked eye where the apFAB51 Measurement Cells look red in ambient light while the Negative Control Cells do not. We are repeating these measurements and using the iGEM recommended protocol for calibrating the fluorescence. We will present the results of the second set of measurements at the iGEM Grand Jamboree.


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]