Team:NAU-CHINA/Composite Part

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Our Best Composite Part:BBa_K2557010

We combined three promoters, miniCMV promoter, EF1α promoter and Ubc promoter, with three recombinases, Bxb1, PhiC31 and TP901, and identified nine combinations. In the end, we chose TetO-miniCMV promoter-Bxb1 as our best composite part.

Function In Our Genetic Circuit

This part can convert the input signal of the upstream TEV into the output signal of the downstream RFP. When TEV is detached from the cell membrane, it functions as a protease, and the inhibition of the promoter by TetR is released, and the expression of Bxb1 recombinase is turned on. Finally, Bxb1 functions as a recombinase to initiate expression of RFP.

Characterization

TetR can effectively repress Tet operator in HEK 293T

Fig. 1 Inhibition of TetR on promoter with TetO sequence
(a)Fluorescence microscope observation of HEK 293T only transfected with plasmids containing promoters with TetO sequence.
(b)Fluorescence microscope observation of HEK 293T transfected with plasmids containing promoters with TetO sequence and tetR.

Bxb1 works well in HEK 293T

Fig. 1a Bxb1 recombinase have a higher fluorescence intensity than those with TP901 recombinase under the same promoter strength and recombinase concentration.
Fig. 1b The reverse efficiency of Bxb1 recombinase is higher than TP901 recombinase under the same promoter strength and recombinase concentration.

Combining these data and, to our knowledge, we first constructed a TetO-regulated recombinase genetic circuit in mammalian cells, so we present BBa_K2557010 as the Our Best Composite Part.

Fig. 1 Bxb1 structure homology-model provided by SWISS-MODEL.

Composite Parts

A composite part is a functional unit of DNA consisting of two or more basic parts assembled together. BBa_I13507 is an example of a composite part, consisting of an RBS, a protein coding region for a red fluorescent protein, and a terminator.

New composite BioBrick devices can be made by combining existing BioBrick Parts (like Inverters, Amplifiers, Smell Generators, Protein Balloon Generators, Senders, Receivers, Actuators, and so on).

Note

This page should list all the composite parts your team has made during your project. You must add all characterization information for your parts on the Registry. You should not put characterization information on this page. Remember judges will only look at the first part in the list for the Best Composite Part award, so put your best part first!

Best Composite Part Special Prize

To be eligible for this award, this part must adhere to Registry sample submission guidelines and have been sent to the Registry of Standard Biological Parts. If you have a part you wish to nominate your team for this special prize, make sure you add your part number to your judging form and delete the box at the top of this page.

Please note: Judges will only look at the first part number you list, so please only enter ONE (1) part number in the judging form for this prize.