Difference between revisions of "Team:BIT-China/Parts"

 
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<body>
 
<body>
  
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    <ul id="left-nav">
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          <ul id="left-nav">
 
         <li>
 
         <li>
 
             <a>PROJECT</a>
 
             <a>PROJECT</a>
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                 <li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsFeedback">Feedback</a></li>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsFeedback">Feedback</a></li>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsOutput">Output</a></li>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsOutput">Output</a></li>
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                 <li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsInput">Input</a></li>
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                 <li><a href="https://2018.igem.org/Team:BIT-China/Results">Results</a></li>
 
             </ul>
 
             </ul>
 
         </li>
 
         </li>
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             <ul>
 
             <ul>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/Model">Overview</a></li>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/Model">Overview</a></li>
−
                 <li><a href="https://2018.igem.org/Team:BIT-China/FluorescentProbesModel">Fluorescent Probes Model </a></li>
+
                 <li><a href="https://2018.igem.org/Team:BIT-China/FluorescentProbesModel">Fluorescent Probe Model </a></li>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/H2O2DecompositionModel">H<sub>2</sub>O<sub>2</sub>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/H2O2DecompositionModel">H<sub>2</sub>O<sub>2</sub>
 
                         Decomposition Model</a></li>
 
                         Decomposition Model</a></li>
  
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/roGFP2-Orp1MichaelisEquationModel">roGFP2-Orp1
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/roGFP2-Orp1MichaelisEquationModel">roGFP2-Orp1
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                         Michaelis equations Model</a></li>
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                         Michaelis equation Model</a></li>
 
             </ul>
 
             </ul>
 
         </li>
 
         </li>
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                                         <a href="http://parts.igem.org/Part:BBa_K2765026" target="_Blank">BBa_K2765026</a>
 
                                         <a href="http://parts.igem.org/Part:BBa_K2765026" target="_Blank">BBa_K2765026</a>
 
                                     </td>
 
                                     </td>
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                                     <td>gal1p-yno1-CYC1T</td>
+
                                     <td>GAL1p-yno1-CYC1t</td>
 
                                     <td>Increase ROS Content </td>
 
                                     <td>Increase ROS Content </td>
 
                                     <td>2651 bp</td>
 
                                     <td>2651 bp</td>
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                                         <a href="http://parts.igem.org/Part:BBa_K2765000" target="_Blank">BBa_K2765000</a>
 
                                         <a href="http://parts.igem.org/Part:BBa_K2765000" target="_Blank">BBa_K2765000</a>
 
                                     </td>
 
                                     </td>
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                                     <td>gal1p</td>
+
                                     <td>GAL1p</td>
 
                                     <td>Turn on the expression of dCas9</td>
 
                                     <td>Turn on the expression of dCas9</td>
 
                                     <td>332 bp</td>
 
                                     <td>332 bp</td>
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                                         <a href="http://parts.igem.org/Part:BBa_K2765001" target="_Blank">BBa_K2765001</a>
 
                                         <a href="http://parts.igem.org/Part:BBa_K2765001" target="_Blank">BBa_K2765001</a>
 
                                     </td>
 
                                     </td>
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                                     <td>sod2p</td>
+
                                     <td>SOD2p</td>
 
                                     <td>Turn on the expression of dCas9</td>
 
                                     <td>Turn on the expression of dCas9</td>
 
                                     <td>526 bp</td>
 
                                     <td>526 bp</td>
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                                         <a href="http://parts.igem.org/Part:BBa_K2765002" target="_Blank">BBa_K2765002</a>
 
                                         <a href="http://parts.igem.org/Part:BBa_K2765002" target="_Blank">BBa_K2765002</a>
 
                                     </td>
 
                                     </td>
−
                                     <td>trr1p</td>
+
                                     <td>TRR1p</td>
 
                                     <td>Turn on the expression of dCas9</td>
 
                                     <td>Turn on the expression of dCas9</td>
 
                                     <td>960 bp</td>
 
                                     <td>960 bp</td>
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                                         <a href="http://parts.igem.org/Part:BBa_K2765003" target="_Blank">BBa_K2765003</a>
 
                                         <a href="http://parts.igem.org/Part:BBa_K2765003" target="_Blank">BBa_K2765003</a>
 
                                     </td>
 
                                     </td>
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                                     <td>trx2p</td>
+
                                     <td>TRX2p</td>
 
                                     <td>Turn on the expression of dCas9</td>
 
                                     <td>Turn on the expression of dCas9</td>
 
                                     <td>283 bp</td>
 
                                     <td>283 bp</td>
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                                         <a href="http://parts.igem.org/Part:BBa_K2765005" target="_Blank">BBa_K2765005</a>
 
                                         <a href="http://parts.igem.org/Part:BBa_K2765005" target="_Blank">BBa_K2765005</a>
 
                                     </td>
 
                                     </td>
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                                     <td>gsh1p</td>
+
                                     <td>GSH1p</td>
 
                                     <td>Turn on the expression of dCas9</td>
 
                                     <td>Turn on the expression of dCas9</td>
 
                                     <td>2027 bp</td>
 
                                     <td>2027 bp</td>
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                                         <a href="http://parts.igem.org/Part:BBa_K2765050" target="_Blank">BBa_K2765050</a>
 
                                         <a href="http://parts.igem.org/Part:BBa_K2765050" target="_Blank">BBa_K2765050</a>
 
                                     </td>
 
                                     </td>
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                                     <td>TEF2p-roGFP2-Orp1-CYC1T</td>
+
                                     <td>TEF2p-roGFP2-Orp1-CYC1t</td>
 
                                     <td>Detect ROS Content</td>
 
                                     <td>Detect ROS Content</td>
 
                                     <td>1965 bp</td>
 
                                     <td>1965 bp</td>
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                                         <a href="http://parts.igem.org/Part:BBa_K2765051" target="_Blank">BBa_K2765051</a>
 
                                         <a href="http://parts.igem.org/Part:BBa_K2765051" target="_Blank">BBa_K2765051</a>
 
                                     </td>
 
                                     </td>
−
                                     <td>TEF1p-roGFP2-Orp1- CYC1T</td>
+
                                     <td>TEF1p-roGFP2-Orp1- CYC1t</td>
 
                                     <td>Detect ROS Content</td>
 
                                     <td>Detect ROS Content</td>
 
                                     <td>1964 bp</td>
 
                                     <td>1964 bp</td>
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                                         <a href="http://parts.igem.org/Part:BBa_K2765052" target="_Blank">BBa_K2765052</a>
 
                                         <a href="http://parts.igem.org/Part:BBa_K2765052" target="_Blank">BBa_K2765052</a>
 
                                     </td>
 
                                     </td>
−
                                     <td>ENO2p-roGFP2-Orp1- CYC1T</td>
+
                                     <td>ENO2p-roGFP2-Orp1- CYC1t</td>
 
                                     <td>Detect ROS Content</td>
 
                                     <td>Detect ROS Content</td>
 
                                     <td>2074 bp</td>
 
                                     <td>2074 bp</td>
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                                     <td>16</td>
 
                                     <td>16</td>
 
                                     <td>
 
                                     <td>
−
                                         <a href="http://parts.igem.org/Part:BBa_K2765051" target="_Blank">BBa_K2765051</a>
+
                                         <a href="http://parts.igem.org/Part:BBa_K2765053" target="_Blank">BBa_K2765053</a>
 
                                     </td>
 
                                     </td>
−
                                     <td>FBA1p-roGFP2-Orp1- CYC1T</td>
+
                                     <td>FBA1p-roGFP2-Orp1- CYC1t</td>
 
                                     <td>Detect ROS Content</td>
 
                                     <td>Detect ROS Content</td>
 
                                     <td>2193 bp</td>
 
                                     <td>2193 bp</td>

Latest revision as of 22:20, 17 October 2018

PARTS

In 2018, BIT-China has submitted 16 parts. We divide our parts into 3 group, Regulator, Feedback and Output. Each group contains some parts with similar function or close relation. Different groups also connect with each other.

Regulator
Increase ROS Content

BBa_K2765026 and BBa_K2765021 is a collection of regulate components including coding protein and expression cassette, which can increase the accumulation of intracellular ROS through overexpressing yon1 induced by galactose.

Improve Tolerance

BBa_K2765030 is 1000bp homologous arm of yca1 gene, which can knock out yca1 gene through homologous recombination so that the yeast's tolerance to H2O2 could be improved.

Feedback
Control ROS Content

BBa_K2765000~BBa_K2765005 is a collection of feedback components that are downstream gene promoter of Yap1. We use them to turn on the expression of dCas9 which can inhibit the expression of yno1 gene. Considering the application of our system in the future, to realize different feedback effect, we verified different promoters, GLR1p, SOD2p, TRR1p, TRX2p and GSH1p, with different transcription level.

Output
Detect ROS content

BBa_K2765040~BBa_K2765053 is a collection of output components including promoters, coding protein and expression cassette, which can express redox-sensitive green fluorescent protein roGFP2-Orp1 and display ROS content in cells by its fluorescence ratio.

According to this measurement principle, the concentration of whole roGFP2-Orp1 can influence the final signal output strength, even detect the same sample, so we verified four promoters, FBA1, TEF2, TEF1 and ENO2, with different transcription level to solve this interfere. Considering the application of our system in the future, we made this collection and wished these parts can be used in different antioxidant detection.

Improve a previous part

We performed codon optimization on team Connell 2017's previous part BBa_K2296006 and obtained our core part BBa_K2296006, which codes for roGFP2-Orp1 protein. Also we got As a result, it is more suitable to our chassis organisms---yeast and has better expression in yeast. Check here to know further information.

Part Form
No. Biobrick Name Description Length
Regulator
1 BBa_K2765021 yno1 ROS Content Increaser 1713 bp
2 BBa_K2765026 GAL1p-yno1-CYC1t Increase ROS Content 2651 bp
3 BBa_K2765030 homologous arm-ura(△yca1) Knock out the yca1 gene 3038 bp
Feedback
4 BBa_K2765000 GAL1p Turn on the expression of dCas9 332 bp
5 BBa_K2765001 SOD2p Turn on the expression of dCas9 526 bp
6 BBa_K2765002 TRR1p Turn on the expression of dCas9 960 bp
7 BBa_K2765003 TRX2p Turn on the expression of dCas9 283 bp
8 BBa_K2765005 GSH1p Turn on the expression of dCas9 2027 bp
Output
9 BBa_K2765040 FBA1p Turn on the expression of roGFP2-Orp1 631 bp
10 BBa_K2765042 ENO2p Turn on the expression of roGFP2-Orp1 512 bp
11 BBa_K2765043 roGFP2-Orp1 ROS Content Detector 1317 bp
12 BBa_K2765045 TEF1p Turn on the expression of roGFP2-Orp1 402 bp
13 BBa_K2765050 TEF2p-roGFP2-Orp1-CYC1t Detect ROS Content 1965 bp
14 BBa_K2765051 TEF1p-roGFP2-Orp1- CYC1t Detect ROS Content 1964 bp
15 BBa_K2765052 ENO2p-roGFP2-Orp1- CYC1t Detect ROS Content 2074 bp
16 BBa_K2765053 FBA1p-roGFP2-Orp1- CYC1t Detect ROS Content 2193 bp

Contact

Institute of Biotransformation and synthetic biosystem School of Chemistry and Chemical Engineering.

Beijing Institute of Technology

100081, Beijing

Email:lichun@bit.edu.cn

Copyright © 2018 BIT-China