Difference between revisions of "Part:BBa K3447008"
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===Usage and Biology===
 
===Usage and Biology===
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In our project, we divided T7 RNA polymerase into two parts, respectively called T7<sub>NT</sub> <partinfo>BBa_K3447008</partinfo> and T7<sub>CT</sub> <partinfo>BBa_K3447007</partinfo> and attached on the nMag and pMag. Under blue light irradiation, these two parts undergo specific blue light dependent heterodimerization and assemble into complete T7 RNA polymerase, which acts on PT7 promoter <partinfo>BBa_K3447005</partinfo>and activates downstream gene expression. And the process is reversible, that is, when blue light is stopped, the dimerized T7 RNA polymerase dedimers and loses its function.<br><br>
+
In our project, we divided T7 RNA polymerase into two parts, respectively called T7<sub>NT</sub> (<partinfo>BBa_K3447008</partinfo>) and T7<sub>CT</sub> (<partinfo>BBa_K3447007</partinfo>) and attached on the nMag and pMag. Under blue light irradiation, these two parts undergo specific blue light dependent heterodimerization and assemble into complete T7 RNA polymerase, which acts on PT7 promoter (<partinfo>BBa_K3447005</partinfo>) and activates downstream gene expression. And the process is reversible, that is, when blue light is stopped, the dimerized T7 RNA polymerase dedimers and loses its function.<br><br>
  
 
[[Image: T7 promoter (strong promoter from T7 bacteriophage).jpg|center|frame|100px|<b>Figure 1. </b> Main principles of the molecular mechanism of T7 promoter]]<br><br>
 
[[Image: T7 promoter (strong promoter from T7 bacteriophage).jpg|center|frame|100px|<b>Figure 1. </b> Main principles of the molecular mechanism of T7 promoter]]<br><br>

Revision as of 09:48, 25 October 2020


xynD, which produces xylanase

T7 polymerase stage 1 with nMag.


Usage and Biology

In our project, we divided T7 RNA polymerase into two parts, respectively called T7NT (BBa_K3447008) and T7CT (BBa_K3447007) and attached on the nMag and pMag. Under blue light irradiation, these two parts undergo specific blue light dependent heterodimerization and assemble into complete T7 RNA polymerase, which acts on PT7 promoter (BBa_K3447005) and activates downstream gene expression. And the process is reversible, that is, when blue light is stopped, the dimerized T7 RNA polymerase dedimers and loses its function.

Figure 1. Main principles of the molecular mechanism of T7 promoter


In our project, we use this part to form a complete T7 RNA polymerase by dimerizing it with T7CT under blue light. (链接104+f09+104+f10+001+b17)

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1918
    Illegal PstI site found at 1928
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1918
    Illegal PstI site found at 1928
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1918
    Illegal PstI site found at 1928
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1918
    Illegal PstI site found at 1928
  • 1000
    COMPATIBLE WITH RFC[1000]

Design

Design Notes

We added some synonymous mutations to avoid part rules.



  • Armin Baumschlager, Stephanie K. Aoki, and Mustafa Khammash, Dynamic Blue Light-Inducible T7 RNA Polymerases (Opto-T7RNAPs) for Precise Spatiotemporal Gene Expression Control, ACS Synth. Biol. 2017, 6, 2157-2167.

References

1. Armin Baumschlager, Stephanie K. Aoki, and Mustafa Khammash, Dynamic Blue Light-Inducible T7 RNA Polymerases (Opto-T7RNAPs) for Precise Spatiotemporal Gene Expression Control, ACS Synth. Biol. 2017, 6, 2157-2167.