Difference between revisions of "Part:BBa K2272000"
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===Usage and Biology===
 
===Usage and Biology===
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[[File:PVHb_data.jpeg|thumb|600px|center|Read-out fluoresence level of GFP from the reporter circuit of pVHb comparing to dissolved oxygen level and cell density.]]
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<p>VhB has been described in literature as being maximally active in hypoxic conditions, ideally at 2% dissolved oxygen saturation When tested with the above describe protocol, results are inconclusive. The first trial was conducted on 9/25 and yielded positive data that corroborated previous evidence of maximal induction under hypoxic conditions. As time spend oped increased, measurements of relative fluorescence decreased, indicating inhibition of GFP expression. There was almost no observed relationship between relative fluorescence and dissolved oxygen content.</p>
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[[File:Virginia_VHb1_David.png|thumb|600px|center|Figure 5: Scatterplots of relative fluorescence versus time and dissolved oxygen content DH5a E. coli transformed with the VhB reporter biobrick.]]
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<p>When tested on 10/22, there is a clear positive relationship between time spent exposed to atmospheric oxygen and the intensity of the relative fluorescence. This data is not only contradictory to the other data collected on 9/25, but it is also contradictory to the results found in literature and by other iGEM teams. Because of this, it is reasonable to conclude that there may be a bias in the measurement of fluorescence emitted by cells expressing the VhB reporter biobrick.</p>
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[[File:Virginia_VHb2_David.png|thumb|600px|center|Figure 6: Scatterplots of relative fluorescence versus time and dissolved oxygen content DH5a E. coli transformed with the VhB reporter biobrick.]]
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2272000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2272000 SequenceAndFeatures</partinfo>

Revision as of 03:19, 1 November 2017


promoter VHb

A promoter that is maximally induced under microaerobic condition and regulates the expression of VHb, a bacterial hemoglobin that acts as a terminal oxidase.


Usage and Biology

VhB has been described in literature as being maximally active in hypoxic conditions, ideally at 2% dissolved oxygen saturation When tested with the above describe protocol, results are inconclusive. The first trial was conducted on 9/25 and yielded positive data that corroborated previous evidence of maximal induction under hypoxic conditions. As time spend oped increased, measurements of relative fluorescence decreased, indicating inhibition of GFP expression. There was almost no observed relationship between relative fluorescence and dissolved oxygen content.

Figure 5: Scatterplots of relative fluorescence versus time and dissolved oxygen content DH5a E. coli transformed with the VhB reporter biobrick.

When tested on 10/22, there is a clear positive relationship between time spent exposed to atmospheric oxygen and the intensity of the relative fluorescence. This data is not only contradictory to the other data collected on 9/25, but it is also contradictory to the results found in literature and by other iGEM teams. Because of this, it is reasonable to conclude that there may be a bias in the measurement of fluorescence emitted by cells expressing the VhB reporter biobrick.

Figure 6: Scatterplots of relative fluorescence versus time and dissolved oxygen content DH5a E. coli transformed with the VhB reporter biobrick.

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]