Difference between revisions of "Part:BBa K4205111:Experience"
| Line 19: | Line 19: | ||
<!-- End of the user review template --> | <!-- End of the user review template --> | ||
<!-- DON'T DELETE --><partinfo>BBa_K4205111 EndReviews</partinfo> | <!-- DON'T DELETE --><partinfo>BBa_K4205111 EndReviews</partinfo> | ||
| + | <p>Since the 3D structure of LldR in our chassis, Escherichia coli Nissle 1917, was not found in PDB, we used SWISS-MODEL for homology modeling to predict the 3D structure of LldR.</p> | ||
| + | <p>With the help of AutoDock, we simulated the docking of LldR and L-lactate. The resulting figure is presented below. The docking sites are colored yellow.</p> | ||
| + | <p>We constructed the ALPaGA-lacZ plasmid and transformed it into ''Escherichia coli'' DH5α. To prove the success of our transformation, we designed primers, performed plasmid PCR and colony PCR to amplify the LldR gene, and ran the agarose gal.</p> | ||
| + | <p>As shown in the figure below, after plasmid PCR and colony PCR, the sample from the transformed DH5α both ran out bands near 750 bp, which was consistent with the target band (777 bp). This further indicated that our plasmid transformation was successful.</p> | ||
| + | [[File:lldR SWISS.png|300px|thumb|left|Predicted 3D structure of LldR]] | ||
| + | [[File:lldr and lactate.png|300px|thumb|left|Docking of LldR and L-lactate]] | ||
| + | [[File:4-LldRPCR.png|700px|thumb|left|Proof of the successful transformation.(A) plasmid PCR. Lanes 1-2: PCR sample form transformed DH5α. Lanes 3-4: PCR sample form untransformed DH5α. | ||
| + | (B) colony PCR. Lanes 1-4: PCR sample form transformed DH5α. Lanes 5-8: PCR sample form untransformed DH5α. | ||
| + | ]] | ||
Revision as of 10:58, 12 October 2022
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K4205111
User Reviews
UNIQa00b6ea862dbfe57-partinfo-00000000-QINU UNIQa00b6ea862dbfe57-partinfo-00000001-QINU
Since the 3D structure of LldR in our chassis, Escherichia coli Nissle 1917, was not found in PDB, we used SWISS-MODEL for homology modeling to predict the 3D structure of LldR.
With the help of AutoDock, we simulated the docking of LldR and L-lactate. The resulting figure is presented below. The docking sites are colored yellow.
We constructed the ALPaGA-lacZ plasmid and transformed it into Escherichia coli DH5α. To prove the success of our transformation, we designed primers, performed plasmid PCR and colony PCR to amplify the LldR gene, and ran the agarose gal.
As shown in the figure below, after plasmid PCR and colony PCR, the sample from the transformed DH5α both ran out bands near 750 bp, which was consistent with the target band (777 bp). This further indicated that our plasmid transformation was successful.
