Difference between revisions of "Team:BIT/Composite Part"

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<h3>★  ALERT! </h3>
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<link href="https://2016.igem.org/Team:BIT/css/bootstrap?action=raw&ctype=text/css"
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<p>This page is used by the judges to evaluate your team for the <a href="https://2016.igem.org/Judging/Awards#Special_Prizes">Composite Part special prize</a>. </p>
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<p> Delete this box in order to be evaluated for this medal. See more information at <a href="https://2016.igem.org/Judging/Pages_for_Awards/Instructions"> Instructions for Pages for awards</a>.</p>
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A composite part is a functional unit of DNA consisting of two or more basic parts assembled together. <a href="http://parts.igem.org/wiki/index.php/Part:BBa_I13507">BBa_I13507</a> is an example of a composite part, consisting of an RBS, a protein coding region for a red fluorescent protein, and a terminator.
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</p>
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<p>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).</p>
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#home{
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background: url(https://static.igem.org/mediawiki/2016/9/97/BIT-BIology.jpg);
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<h4>Note</h4>
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<p>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.</p>
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<h1><font color=#c91f77>C</font>OMPOSITE <font color=#c91f77>P</font>ART</h1>
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    <li class=color1><a href="https://2016.igem.org/Team:BIT/Composite_Part"><h2>Composite Part</h2></a></li>
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<h2><br><br><br><br><br><br><br>Composite Part</h2>
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<center> <font size="1px">ptet-RBS-TetR-FimE-T-pbad-BxB1-T-constitutive promoter-two integrase sites-RBS-GFP-T</font></center><br><br>
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</div>
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<p>Integrase is able to identify a different specific sequences, called attP (on phage) and attB (in the host). After restructured and inserted in the phage, attP and attB will be flipped and converted into a pair of sites called attL and attR, which are no longer substrates for the recombinase. Using attP and attB sites, deliberately construct a modified DNA sequence, in this way you can "cheat" recombinase, so that the DNA sequence occur to irreversible reversal. The part contains the FimE integrase as well as its recognition sites attP, attB and BxB1 integrase as well as its recognition sites IRR, IRL. We use FimE and BxB1 to control the generation of GFP. In addition, the inhibition mechanisms of tetR protein on ptet promoter and LacI protein on plac promoter are used to control the expression of FimE and BxB1. If to add a certain amount of tetracycline to the inoculum, tetracycline will combine with TetR protein, the inhibition of TetR on ptet will disappear, and the downstream genes of FimE and BxB1 integrase can express. They will respectively act on the recognition sites behind the constitutive promoter, so that the site occurs 180 ° reversal and the original positive terminator becomes the reverse terminator, with the loss of the role of termination of transcription. Finally, the GFP can be expressed. This system can verify the function of integrase, also can be used as a detection mechanism to detect the presence of tetracycline, thus providing the theoretical basis for the development of our formal experiment. Using this mechanism can detect more material by changed the promoter and the promoter’s inhibitors. </p>
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Haidian Beijing 100081, P. R. China<br><br>
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Latest revision as of 03:43, 20 October 2016

<!DOCTYPE html> Biology

PARTS

COMPOSITE PART








Composite Part


...
ptet-RBS-TetR-FimE-T-pbad-BxB1-T-constitutive promoter-two integrase sites-RBS-GFP-T


Integrase is able to identify a different specific sequences, called attP (on phage) and attB (in the host). After restructured and inserted in the phage, attP and attB will be flipped and converted into a pair of sites called attL and attR, which are no longer substrates for the recombinase. Using attP and attB sites, deliberately construct a modified DNA sequence, in this way you can "cheat" recombinase, so that the DNA sequence occur to irreversible reversal. The part contains the FimE integrase as well as its recognition sites attP, attB and BxB1 integrase as well as its recognition sites IRR, IRL. We use FimE and BxB1 to control the generation of GFP. In addition, the inhibition mechanisms of tetR protein on ptet promoter and LacI protein on plac promoter are used to control the expression of FimE and BxB1. If to add a certain amount of tetracycline to the inoculum, tetracycline will combine with TetR protein, the inhibition of TetR on ptet will disappear, and the downstream genes of FimE and BxB1 integrase can express. They will respectively act on the recognition sites behind the constitutive promoter, so that the site occurs 180 ° reversal and the original positive terminator becomes the reverse terminator, with the loss of the role of termination of transcription. Finally, the GFP can be expressed. This system can verify the function of integrase, also can be used as a detection mechanism to detect the presence of tetracycline, thus providing the theoretical basis for the development of our formal experiment. Using this mechanism can detect more material by changed the promoter and the promoter’s inhibitors.






Contact Us

Address

Beijing Institute of Technology,
No. 5 South Zhong Guan Cun Street,
Haidian Beijing 100081, P. R. China

Twitter : @igem_BIT

Sina Weibo : @igem_BIT

Website : http://www.bit.edu.cn
Send