Difference between revisions of "Part:BBa K3596030"
Line 15: Line 15:
 
</p>
 
</p>
 
<div style="text-align: center;">
 
<div style="text-align: center;">
−
<img src="https://2020.igem.org/wiki/images/4/4e/T--GreatBay_SZ--western.png" alt="" width="700">
+
<img src="https://2020.igem.org/wiki/images/4/4e/T--GreatBay_SZ--western.png" alt="" width="400">
 
<h6 style="text-align:center">Figure 2: Western Blot of the cell lysis of BIOT and BIOT-His(express e-PN and e-PN-His respectively); Antibody: Anti-His tag; e-PN-His: 7.7 kDa.</h6>   
 
<h6 style="text-align:center">Figure 2: Western Blot of the cell lysis of BIOT and BIOT-His(express e-PN and e-PN-His respectively); Antibody: Anti-His tag; e-PN-His: 7.7 kDa.</h6>   
  

Revision as of 04:44, 25 October 2020

BIOT: PpdD Signal+Pilin Gene+ TypeIV pilin assembly system

BIOT is a basic design for the expression of Geobacter sulfurreducens' 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

The e-PN can be successfully expressed by express this part under the control of pTac, and the result was verified by Western Blot(figure 2).

Figure 2: Western Blot of the cell lysis of BIOT and BIOT-His(express e-PN and e-PN-His respectively); Antibody: Anti-His tag; e-PN-His: 7.7 kDa.

We made a protein film following the instructions from the paper we researched.[1] And we eventually succeeded in producing stable voltage at about 0.35V with a current of 500nA or so.(figure 3).

Figure 3: The voltage and current of e-PN

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 NgoMIV site found at 6145
    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