Difference between revisions of "Part:BBa K3977002"

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=Usage and Biology=
 
=Usage and Biology=
 
==Characterized by CAFA_China 2022==
 
==Characterized by CAFA_China 2022==
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*We conducted a simple test to see if our design met the expection.
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*The mScarlet-I gene and the relE toxin gene were fused to construct a fusion gene, in order to detect the amount of relE protein synthesis. We removed the terminator of relE and fused it to the mScarlet-I gene.  
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*We designed the genetic circuit that contains sulAp ([[Part:BBa_K518010|sulA promoter]]), Amp30E, relE gene ([[Part:BBa_K185047|RelE toxin]]), mScarlet-I and 3WJ-Bro([[Part:BBa K4226000 |3WJ-Bro]]) , and transferred it into DH10B competent cells (The control group did not contain the Amp30E Amplification Device).  
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*We designed the genetic circuit that contains sulAp ([[Part:BBa_K518010|sulA promoter]]), Amp30E, relE gene ([[Part:BBa_K185047|RelE toxin]]), mScarlet-I and 3WJ-Bro([[Part:BBa K4226000 |3WJ-Bro]]) , and transferred it into DH10B competent cells (The control group did not contain the Amp30E Amplification Device). The diagram is shown bellow:
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*The relE toxin gene and the mScarlet-I gene were fused to construct a fusion gene, in order to detect the amount of relE protein synthesis. We removed the terminator of relE and fused it to the mScarlet-I gene.  
+
 
 +
[[File:PSB1C3-sulAp-Amp30E-relE-mScarlet-I-3WJ-Bro.png|600px|thumb|center|Gene circuit: pSB1C3-sulAp-Amp30E-relE-mScarlet-I-3WJ-Bro]]<br>
 +
 
 +
*After bacterial culture and UV induction, we measured the OD600 and fluorescence values under 579/616nm. Then, the fluorescence per OD was calculated.  
 +
*The curve of experimental group was higher than that of control group as shown in the following figure, indicating the mScarlet-I correctly displayed the increasing tendency of relE protein synthesis under the effect of Amp30E Amplification Device.
 +
 
 +
[[File:MScarlet-I-2022.png|600px|thumb|center|Figure: The mScarlet-I gene was fused with relE gene to detect the amount of relE protein synthesis (579/616nm). Control group: pSB1C3-sulAp-relE-mSarlet-l-3WJxBro; experimental group: pSB1C3-sulAp-Amp30E-relE-mSarlet-l-3WJxBro.]]<br>
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Latest revision as of 08:35, 26 September 2022


mSarlet-I

mSarlet-I

Usage and Biology

Characterized by CAFA_China 2022

  • The mScarlet-I gene and the relE toxin gene were fused to construct a fusion gene, in order to detect the amount of relE protein synthesis. We removed the terminator of relE and fused it to the mScarlet-I gene.
  • We designed the genetic circuit that contains sulAp (sulA promoter), Amp30E, relE gene (RelE toxin), mScarlet-I and 3WJ-Bro(3WJ-Bro) , and transferred it into DH10B competent cells (The control group did not contain the Amp30E Amplification Device). The diagram is shown bellow:
Gene circuit: pSB1C3-sulAp-Amp30E-relE-mScarlet-I-3WJ-Bro

  • After bacterial culture and UV induction, we measured the OD600 and fluorescence values under 579/616nm. Then, the fluorescence per OD was calculated.
  • The curve of experimental group was higher than that of control group as shown in the following figure, indicating the mScarlet-I correctly displayed the increasing tendency of relE protein synthesis under the effect of Amp30E Amplification Device.
Figure: The mScarlet-I gene was fused with relE gene to detect the amount of relE protein synthesis (579/616nm). Control group: pSB1C3-sulAp-relE-mSarlet-l-3WJxBro; experimental group: pSB1C3-sulAp-Amp30E-relE-mSarlet-l-3WJxBro.


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 667
  • 1000
    COMPATIBLE WITH RFC[1000]