Part:BBa_K2336035
oprF-GS-Sitag-FLAG
The capture 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. FLAG is designed to test fluorescence. The genetic engineering bacteria can express the protein on its cell membrane. Then the bacteria can combine with silica surface. So we can get them back. Figure 1 shows the circuit of it.
As we’ve mentioned, this is the final step. After sensing and capturing task finished, the bacteria will combine with silica net, so we can achieve our goal -- get them back (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 (https://parts.igem.org/Part:BBa_K1592007). Two years ago, we did it in yeast. Now we put gene into E.coli. It was a hard experiment, but finally we made it. The gene order is also different from that in eukaryotes, such as yeast. We consider it as a great 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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