Difference between revisions of "Team:NAU-CHINA/Basic Part"

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    <title>Safety</title>
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            <h1>Our Best Basic Part:BBa_K2557001</h1>
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            <p>This year, our team has selected a series of recombinases through literature searches and characterized a number of them. Here we present the best characterized recombinase - Bxb1 (BBa_BBa_K2557001) as our best basic part for the award.</p>
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                <h2>Background</h2>
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                <p>Bxb1, also known as Bxb1 gp35, is an recombinase comeing from Mycobacterium Phage Bxb1 and can bind to specific attB/P sites to catalyze DNA recombination. It helps the phage to integrate its genome into bacterial genome naturally. By constructing the attB/P sites in different directions, Bxb1 gp35 can catalyze the recombination of DNA between their sites, leading to inversion when attB/P are in opposite directions and excision when attB/P are in the same directions. The resulting attL and attR sequences cannot be recognized and bound by integrases alone, so the state after integration is stable. Bxb1 gp35 is widely used to construct combinational logic gate and synthetic biology.</p>
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                    <figcaption class="_table">Fig. 1 Bxb1 structure homology-model provided by SWISS-MODEL.</figcaption>
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                <h2>Function In our Genetic Circuit</h2>
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                <p>In our genetic circuit, Bxb1 regulated by TetR. When the inhibition released by TEV protease, Bxb1 can bind to specific attB/P sites to catalyze DNA recombination, leading to the expression of RFP.</p>
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                <h2>Characterization</h2>
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                <p>Method: Co-transfected with six different numbers of plasmids containing recombinase genes(miniCMV-Bxb1 and miniCMV-TP901) and fixed numbers of plasmids containing corresponding recombinase recognition sites. After 36 hours of plasmid co - transfection, the proportion of fluorescent cells and the average fluorescence intensity of cells were detected by flow cytometry. The experiment was repeated three times.</p>
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                <p>Result:</p>
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                    <figcaption class="_table">Fig. 2a Bxb1 recombinase have a higher fluorescence intensity than those with TP901 recombinase under the same promoter strength and recombinase concentration.</figcaption>
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                    <figcaption class="_table">Fig. 2b The reverse efficiency of Bxb1 recombinase is higher than TP901 recombinase under the same promoter strength and recombinase concentration.
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                <p>Bxb1 works well in HEK 293T as expected, so we present Bxb1(BBa_K2557001) as our best basic part.</p>
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Revision as of 07:36, 16 October 2018

Template:2018_NAU-CHINA

header
Safety

Our Best Basic Part:BBa_K2557001

This year, our team has selected a series of recombinases through literature searches and characterized a number of them. Here we present the best characterized recombinase - Bxb1 (BBa_BBa_K2557001) as our best basic part for the award.

Background

Bxb1, also known as Bxb1 gp35, is an recombinase comeing from Mycobacterium Phage Bxb1 and can bind to specific attB/P sites to catalyze DNA recombination. It helps the phage to integrate its genome into bacterial genome naturally. By constructing the attB/P sites in different directions, Bxb1 gp35 can catalyze the recombination of DNA between their sites, leading to inversion when attB/P are in opposite directions and excision when attB/P are in the same directions. The resulting attL and attR sequences cannot be recognized and bound by integrases alone, so the state after integration is stable. Bxb1 gp35 is widely used to construct combinational logic gate and synthetic biology.

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

Function In our Genetic Circuit

In our genetic circuit, Bxb1 regulated by TetR. When the inhibition released by TEV protease, Bxb1 can bind to specific attB/P sites to catalyze DNA recombination, leading to the expression of RFP.

Characterization

Method: Co-transfected with six different numbers of plasmids containing recombinase genes(miniCMV-Bxb1 and miniCMV-TP901) and fixed numbers of plasmids containing corresponding recombinase recognition sites. After 36 hours of plasmid co - transfection, the proportion of fluorescent cells and the average fluorescence intensity of cells were detected by flow cytometry. The experiment was repeated three times.

Result:

Fig. 2a Bxb1 recombinase have a higher fluorescence intensity than those with TP901 recombinase under the same promoter strength and recombinase concentration.
Fig. 2b The reverse efficiency of Bxb1 recombinase is higher than TP901 recombinase under the same promoter strength and recombinase concentration.

Bxb1 works well in HEK 293T as expected, so we present Bxb1(BBa_K2557001) as our best basic part.

Basic Parts

A basic part is a functional unit of DNA that cannot be subdivided into smaller component parts. BBa_R0051 is an example of a basic part, a promoter regulated by lambda cl.

Most genetically-encoded functions have not yet been converted to BioBrick parts. Thus, there are many opportunities to find new, cool, and important genetically encoded functions, and refine and convert the DNA encoding these functions into BioBrick standard biological parts.

Note

This page should list all the basic 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 Basic Part award, so put your best part first!

Best Basic 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.