Part:BBa_K2336035
oprF-GS-Sitag-FLAG
The report part of our circuit. It can be expressed to bind silicon
Usage and Biology
oprF is motif cell wall anchor domain protein. GS stands for GS linker. Sitag-FLAG is designed to express specially protein. The genetic engineering bacteria can produce the protein on its membrane. Then the bacteria can be absorbed by silica surface. So we can capture them. Figure 1 shows the circuit of it.
As we have said, this part is a final part. After sensing and capture finish, the bacteria will be bound onto silica surface. We can achieve our goal to get them(with lanthanum). Figure 2 shows how it works.
Characterization
We did some experiment to prove it works.
SDS-PAGE
After oprF-GS-Sitag-FLAG is expressed successfully,we centrifuged the bacteria liquid and separated different proteins by SDS-PAGE.
Figure 3 shows an obvious ~57kDa protein bands of oprF-Sitag in test lane, which cannot be found in control lane. This result proves that the bacteria could express oprF-Sitag successfully.
improvement
This part is an expansion of our team's work in 2015(<a href="https://parts.igem.org/Part:BBa_K1592007">BBa_K1592007</a>). Two years ago, we did it in yeast. Now we put gene into E.coli. It's a hard experiment and we finally make it. The gene order is also different from that in eukaryotes, yeast for instance. We consider it as a big breakthrough.
Next we will do experiment to test its binding ability. We hope it will have the effect as we expected.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 72
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 639
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 3
Illegal AgeI site found at 1527 - 1000COMPATIBLE WITH RFC[1000]
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