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Team:NYMU-Taipei/results/optomagnetic-design1
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We now follow the steps we construct our design to examine and illustrate what we have done so far. | We now follow the steps we construct our design to examine and illustrate what we have done so far. | ||
<font color=green> The total parts we construct and their detailed information will be performed and recorded in our part registry.</font> | <font color=green> The total parts we construct and their detailed information will be performed and recorded in our part registry.</font> | ||
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| + | [[Image:total_NYMU.png|frame|none|Fig. 1|This is the total constructs of our optomagnetic design, the notation for each columns with different colors can be seen in the following paragraphs.]] | ||
As soon as we have the AMB-1 bacteria, first, we do the AMB-1 colony PCR to get the Mms13 protein's DNA sequences as our parts to do the following steps. | As soon as we have the AMB-1 bacteria, first, we do the AMB-1 colony PCR to get the Mms13 protein's DNA sequences as our parts to do the following steps. | ||
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The correcterized gel electrophoresis result is shown below. The detail Mms13's information, link [http://partsregistry.org/wiki/index.php?title=Part:BBa_K624005]. | The correcterized gel electrophoresis result is shown below. The detail Mms13's information, link [http://partsregistry.org/wiki/index.php?title=Part:BBa_K624005]. | ||
| - | The correct and check | + | Then we can do the next step of our construct.(See Figure 1) |
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| + | [[Image:y-13-r_NYMU.png|frame|none|Fig. 1|We use recombinant PCR to assemble (1)mms13-rLuc； (2)YN-mms13-rLuc； (3)Y-mms13-rLuc]] | ||
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| + | , The correct and check | ||
[[Image: c1_NYMU.png|frame|none|Fig. 3: The main design construct. Ori is the origin of replication of AMB-1. Pmsp1 is the promoter for AMB-1.]] | [[Image: c1_NYMU.png|frame|none|Fig. 3: The main design construct. Ori is the origin of replication of AMB-1. Pmsp1 is the promoter for AMB-1.]] | ||
Revision as of 02:19, 6 October 2011
Six Constructs and Experimental Results
We now follow the steps we construct our design to examine and illustrate what we have done so far. The total parts we construct and their detailed information will be performed and recorded in our part registry.
This is the total constructs of our optomagnetic design, the notation for each columns with different colors can be seen in the following paragraphs.
As soon as we have the AMB-1 bacteria, first, we do the AMB-1 colony PCR to get the Mms13 protein's DNA sequences as our parts to do the following steps.
The correcterized gel electrophoresis result is shown below. The detail Mms13's information, link [http://partsregistry.org/wiki/index.php?title=Part:BBa_K624005].
Then we can do the next step of our construct.(See Figure 1)
We use recombinant PCR to assemble (1)mms13-rLuc； (2)YN-mms13-rLuc； (3)Y-mms13-rLuc
, The correct and check
Fig. 3: The main design construct. Ori is the origin of replication of AMB-1. Pmsp1 is the promoter for AMB-1.
Fig. 4: A construct without YC on the CHAMP is our negative control in our design. It would not light up no matter there is magnetic field applied or not.
Fig. 5:. We design this construct with the whole YFP on the N-terminus of Mms13 and the CHAMP design to induce the BRET phenomenon. We suppose that with the CHMAP design, the construct will light up only when magnetic force is applied.
Fig. 6: This design is constructed with the whole YFP on the N-terminus of Mms13 but “without” the CHAMP design. The construct should be a positive control with constant light no matter whether force is applied or not.
Fig. 7: The design construct is similar with construct in Fig. 5. Why we construct this design with the whole YFP on the C-terminus of the CHAMP design instead of Mms13’s N-terminus is that we try to avoid the interferences of DNA transformation with sequences added in front of the natural construct. The construct is supposed to light up when force pull the Mms13 and turn off if no magnetic field is applied.
Fig. 8: In this last construct, only r-luciferase anchored on Mms13’s C-terminus. It will light up in yellow light (with 580 nm wavelength) due to the luciferase reaction, instead of the green light induced from BRET phenomenon (540 nm wavelength).
