Difference between revisions of "Part:BBa K3596056"

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===Characterization===
 
===Characterization===
 
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<p>In order to improve the protein nanowire, we mutated the RBSs of the TypeIV pilin assembly system and the <i>pilA</i> gene to enhance the productivity and output voltage, respectively. This part shows the best combination effect, with the optimum RBS mutation RBS4 which increased the protein productivity by 34.1%(figure 2), and the optimum pilA mutation 4A which increased the electricity productivity by 46%(figure 3).
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<p>In order to improve the protein nanowire, we mutated the RBSs of the TypeIV pilin assembly system and the <i>pilA</i> gene to enhance the productivity and output voltage, respectively. This part shows the best combination effect, with the optimum RBS mutation <a href="https://parts.igem.org/Part:BBa_K3596051">RBS4</a> which increased the protein productivity by 34.1%(figure 2), and the optimum <i>pilA</i> mutation <a href="https://parts.igem.org/Part:BBa_K3596043">4A</a> which increased the electricity productivity by 46%(figure 3).
 
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Revision as of 08:06, 25 October 2020


BIOT with RBS mut4 and Pilin gene mut4A

BIOT is a basic design for the expression of Geobacter sulfurreducens's pilin-based electrically conductive protein nanowires (e-PNs)[1]. The system includes a tac promoter which promotes the pilin gene(pilA) to express protein monomers; the monomers are later assembled into protein nanowires by the TypeIV pilin assembly system, which is constructed from the gene cluster at the downstream of the pilin gene(figure 1). The e-PNs can generate electric potential with the presence of water molecules in the air, which can be further modified for other purposes.

Figure 1: A Construction of BIOT gene circuit

Characterization

In order to improve the protein nanowire, we mutated the RBSs of the TypeIV pilin assembly system and the pilA gene to enhance the productivity and output voltage, respectively. This part shows the best combination effect, with the optimum RBS mutation RBS4 which increased the protein productivity by 34.1%(figure 2), and the optimum pilA mutation 4A which increased the electricity productivity by 46%(figure 3).

Figure 2: Different pilin productivity of engineered BIOT system with different RBS.
Figure 3: Different electricity productivity of engineered BIOT system with different pilin mutant.

References

[1] Liu X, Gao H, Ward J E, et al. Power generation from ambient humidity using protein nanowires[J]. Nature, 2020, 578(7796): 550-554.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2393
    Illegal BglII site found at 2857
    Illegal BglII site found at 4658
    Illegal BamHI site found at 1073
    Illegal BamHI site found at 3182
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1300
    Illegal AgeI site found at 1146
    Illegal AgeI site found at 1439
    Illegal AgeI site found at 2002
    Illegal AgeI site found at 4626
    Illegal AgeI site found at 6914
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 7920
    Illegal SapI.rc site found at 4262