Difference between revisions of "Part:BBa K2012015"

 
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PcpcG2 promoter is a 238bp green-light activated promoter in Synechocystis PCC 6803(Part:BBa_K592003). The full length promoter is comprised of a G-box region, a CcaR-P activated promoter, and a constitutive promoter, which contributes to the leakiness under red light and low dynamic range. Therefore, we constructed PcpcG2-172, a 172bp truncated cpcG2 promoter deleted for the constitutive promoter.
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PcpcG2 promoter is a 238bp green-light activated promoter in Synechocystis PCC 6803(Part:BBa_K592003). The full length promoter is comprised of a G-box region, a CcaR-P activated promoter, and a constitutive promoter, which contributes to the leakiness under red light and low dynamic range. Therefore, we constructed PcpcG2-172, a 172bp truncated cpcG2 promoter deleted for the constitutive promoter.(For more detail, please  view our wiki: "http://2016.igem.org/Team:HZAU-China/Experiments" )
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<div style="margin-left:auto;margin-right:auto;position:relative"><p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:宋体">世界名校情况<span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:宋体">获奖比例<span></span></span></p>
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<div style="margin-left:auto;margin-right:auto;position:relative"><p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"><img height="318.92999267578125" src="https://static.igem.org/mediawiki/2016/5/5f/Eeeee.jpeg" width="480.0"></span><span style="font-family:Calibri"><span></span></span></p>
 
<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri">&nbsp;</span></p>
 
<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri">&nbsp;</span></p>
<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri">&nbsp;</span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri">iGEM</span><span style="color:#333333;font-family:宋体">竞赛由美国麻省理工学院 2003 年创办,2005年发展成为国际赛事,近来来发展迅猛、规模不断扩大。</span><span style="font-family:宋体">每年秋季在MIT, Boston举办Giant Jamboree,蜂拥而至的合成生物学爱好者来自世界各地名校,包括麻省理工学院、哈佛大学、耶鲁大学、剑桥大学、牛津大学、斯坦福大学、帝国理工学院、海德堡大学、加州大学伯克利分校等世界顶尖学府的代表队。2015年参赛队伍有282支,其中金奖115枚,银奖57,铜奖55. 2014年303支队伍中99金牌,36银牌,59铜牌。今年参赛队伍306支,将在10月底赴美比赛。<span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"><strong>Figure 1.</strong> Fluorescence assay of CcaS-CcaR system with PcpcG2-172 (BBa_K2012015) in CL1 (△EnvZ), and PCB (△CcaS) as </span><span style="font-family:Calibri">chromophore</span><span style="font-family:Calibri">. </span><span style="font-family:Calibri"><span></span></span></p>
<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri"><img height="169.60000610351562" src="http://b200.photo.store.qq.com/psb?/V10XRB7b0MY7nJ/GKxIDufDrn0JKbxQYFeMdFMCvtj23W3RwyrsjN585qI!/b/dMgAAAAAAAAA&amp;bo=.QCqAAAAAAADAHY! " width="248.52999877929688"></span><span style="font-family:Calibri"><span>&nbsp;</span></span><span style="font-family:Calibri"><img height="170.6699981689453" src="http://b262.photo.store.qq.com/psb?/V10XRB7b0MY7nJ/k38U4m2580dGZN..BvcCMdnJkGAjnrurJj.a8*KLNgY!/b/dAYBAAAAAAAA&amp;bo=DgGrAAAAAAADAIE! " width="269.8699951171875"></span><a name="_GoBack"></a><span style="font-family:Calibri"><span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri">F</span><span style="font-family:Calibri">luorescence under green light is 1.81-folds of red light, proving that green light activates output expression, the device works well. </span><span style="font-family:Calibri">PcpcG2-172 </span><span style="font-family:Calibri">shows high efficiency.<span></span></span></p>
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<div style="margin-left:auto;margin-right:auto;position:relative"><p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">The mechanism of bio-pattern formation has been studied for nearly half a century, and many theories have been proposed. After entering the 20th century, with the development of Synthetic Biology, more and more biologist start to research it with the ideology of Synthetic, and significant achievements have been achieved.<span>&nbsp; </span>recent years, with the rapid development of computer science, computers have entered almost every household, and became a very important part of works and entertainments in our life. Now, Augmented reality(AR) gradually walk into people's horizons and Man-Computer Interaction bring us new visual stimulation and life feelings. For introducing AR to the study of bio-pattern formation. We<span>&nbsp; </span>use<span>&nbsp; </span>light sense bacterials to make it's movability can be controlled by gene circuits. Then, we created a pattern we wanted in computer and compare it with the shape of the lawn<span>&nbsp; </span>we collect through CCD. The result of compare will be transformed and used to control the move of part bacterial in the colony to change the shape of colony, and it will be the shape in computer finally. Using this technology, we will control the process of bio-tissue development and make organ regeneration precisely possible.<span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">&nbsp;</span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">B</span><span style="font-family:Calibri;font-size:12.0pt">io-pattern formation </span><span style="font-family:Calibri;font-size:12.0pt"><span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:宋体;font-size:12.0pt">用synthetic研究bpf</span><span style="font-family:Calibri;font-size:12.0pt"><span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">P</span><span style="font-family:Calibri;font-size:12.0pt">C</span><span style="font-family:Calibri;font-size:12.0pt"><span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">AR</span><span style="font-family:Calibri;font-size:12.0pt"><span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:宋体;font-size:12.0pt">光控制基因回路,然后用计算机比较,然后改菌落形态,得到想要的图案</span><span style="font-family:Calibri;font-size:12.0pt"><span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:宋体;font-size:12.0pt">应用:再生医学</span><span style="font-family:Calibri;font-size:12.0pt"><span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">&nbsp;</span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">What the Hell!</span><span style="font-family:Calibri;font-size:12.0pt"><span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">&nbsp;</span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">&nbsp;</span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">Have you ever wondered how the stripes on a clownfish come into being? Bio-pattern formation is formation of spatial pattern of a group of cells in morphogenesis. When guided by computer, the spatial pattern of a group of cells can be altered on request, thus establishing bio-pattern formation </span><span style="font-family:Calibri;font-size:12.0pt">in Augmented </span><span style="font-family:Calibri;font-size:12.0pt">R</span><span style="font-family:Calibri;font-size:12.0pt">eality</span><span style="font-family:Calibri;font-size:12.0pt"> </span><span style="font-family:Calibri;font-size:12.0pt">(AR)</span><span style="font-family:Calibri;font-size:12.0pt">. In our study, a light-switchable E. coli is presented to </span><span style="font-family:Calibri;font-size:12.0pt">control synthetic gene circuits</span><span style="font-family:Calibri;font-size:12.0pt"> including the mobility of cell. </span><span style="font-family:Calibri;font-size:12.0pt">B</span><span style="font-family:Calibri;font-size:12.0pt">y comparing the input pattern with the </span><span style="font-family:Calibri;font-size:12.0pt">shape of the lawn</span><span style="font-family:Calibri;font-size:12.0pt"> in reality</span><span style="font-family:Calibri;font-size:12.0pt"> collect</span><span style="font-family:Calibri;font-size:12.0pt">ed</span><span style="font-family:Calibri;font-size:12.0pt"> through CCD</span><span style="font-family:Calibri;font-size:12.0pt">, </span><span style="font-family:Calibri;font-size:12.0pt">a </span><span style="font-family:Calibri;font-size:12.0pt">light induced </span><span style="font-family:Calibri;font-size:12.0pt">circuit </span><span style="font-family:Calibri;font-size:12.0pt">is </span><span style="font-family:Calibri;font-size:12.0pt">generate</span><span style="font-family:Calibri;font-size:12.0pt">s</span><span style="font-family:Calibri;font-size:12.0pt"> </span><span style="font-family:Calibri;font-size:12.0pt">by the computer, </span><span style="font-family:Calibri;font-size:12.0pt">involving regulatory feedback to control the </span><span style="font-family:Calibri;font-size:12.0pt">mobility of every bacteria. The morphogenetic fate of each cell is then re-determined, which leads to the transformation of the lawn. Eventually, the shape of the lawn matches the primary input pattern in the computer identically. Utilizing this </span><span style="font-family:Calibri;font-size:12.0pt">unparalleled</span><span style="font-family:Calibri;font-size:12.0pt"> technology, </span><span style="font-family:Calibri;font-size:12.0pt">the process of bio-tissue development </span><span style="font-family:Calibri;font-size:12.0pt">can be manipulated precisely, not to mention</span><span style="font-family:Calibri;font-size:12.0pt"> </span><span style="font-family:Calibri;font-size:12.0pt">the feasibility of </span><span style="font-family:Calibri;font-size:12.0pt">organ regeneration </span><span style="font-family:Calibri;font-size:12.0pt">in medical application.</span><span style="font-family:Calibri;font-size:12.0pt"><span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">&nbsp;</span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:宋体;font-size:12.0pt">黄师傅</span><span style="font-family:Calibri;font-size:12.0pt"><span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">our project is based on the concept of augmented reality.the purpose of our project is to<span>&nbsp; </span>stimulate the enviroment of Ec oli<span>&nbsp; </span>on a Lb PLATE just like the pokemon go.to approve this concept we choose to retrieve its chemotaxis with light.while processing our experiment we find it really hard to concisely regulate the bacteria by light,then we decide to turn to a new tool,the triple-tandem riboswitch to help us concisely augment the reality of bacteria.the application of our idea is beyond imagination,not only chemotaxis,we can also stimulate other enviroments on the surface of a plate like the invasion of phage without interact with real virus which can help improve the safty of biological experiment.ofcourse it also has many other applications.<span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">&nbsp;</span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">Terminator!!!<span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">Bio-pattern formation is the establishment of spatial patterns in morphogenesis. When guided by computer, the spatial pattern of a group of cells can be altered in an augmented reality space created </span><a name="_GoBack"></a><span><span style="font-family:Calibri;font-size:12.0pt">by superposing a computer simulated virtual space on a real culture apparatus.</span></span><span></span><span style="font-family:Calibri;font-size:12.0pt"> In our project, light-switchable synthetic gene circuits are adopted to control the mobility of E. coli cells by light-mediated real-time communication between the real bio-pattern in culture media and its virtual counterpart in computer simulation to implement a synchronized growth. Eventually, the shape of the colony matches the preset pattern we want and thus a “what you see is what you get” platform for the study of bio-pattern formation is obtained. Our system can also be extended to eukaryotic cell community to modulate cell fates in the future which will be particularly useful in aiding the development of biological tissues and the regeneration of organs.<span></span></span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:Calibri;font-size:12.0pt">&nbsp;</span></p>
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<p style="text-align:justify;text-justify:inter-ideograph"><span style="font-family:宋体;font-size:12.0pt">生物模式形成是形态建成中空间格局的建立过程。在计算机的指导作用下,通过在现实的培养皿上叠加一个计算机模拟的虚拟空间,我们可以改变增强现实空间中的细胞群体的空间模式。在我们的项目中,采用光控合成基因回路以控制大肠杆菌细胞的运动性。这里通过现实培养的生物模式与其在计算机模拟中的对应部分之间的光控实时交流,使两者实现同步生长。最终,菌落的形状与我们预设的图案完全吻合,因此建立“所见即所得”的生物模式形成的研究平台。我们的系统在未来可以衍生到真核细胞群落,调节它们的细胞命运对于生物组织发育和器官再生有相当大的帮助。</span><span style="font-family:Calibri;font-size:12.0pt"><span></span></span></p>
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<b>Improvement</b><br>A truncated promoter PcpcG2-68 has the similar pattern under the control of green/red light as PcpcG2-172, but is shorter and more convenient to construct. For example, when constructing an array of sgRNAs, PcpcG2-68 has a big advantage over PcpcG2-172: reducing the volume of the circuit and cheaper to synthesis.<br> [[File:T--XHD-Wuhan-B-China--111(1).png|900px|thumb|left|]]<br>Please view https://parts.igem.org/Part:BBa_K3921000 for more details.
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<span id="IGEM 2021 XHD- Wuhan-B-China"><br><br><br><br><br></span>

Latest revision as of 06:41, 21 October 2021


PcpcG2-172, a modified PcpcG2 promoter

PcpcG2 promoter is a 238bp green-light activated promoter in Synechocystis PCC 6803(Part:BBa_K592003). The full length promoter is comprised of a G-box region, a CcaR-P activated promoter, and a constitutive promoter, which contributes to the leakiness under red light and low dynamic range. Therefore, we constructed PcpcG2-172, a 172bp truncated cpcG2 promoter deleted for the constitutive promoter.(For more detail, please view our wiki: "http://2016.igem.org/Team:HZAU-China/Experiments" )

 

Figure 1. Fluorescence assay of CcaS-CcaR system with PcpcG2-172 (BBa_K2012015) in CL1 (△EnvZ), and PCB (△CcaS) as chromophore.

Fluorescence under green light is 1.81-folds of red light, proving that green light activates output expression, the device works well. PcpcG2-172 shows high efficiency.




Improvement
A truncated promoter PcpcG2-68 has the similar pattern under the control of green/red light as PcpcG2-172, but is shorter and more convenient to construct. For example, when constructing an array of sgRNAs, PcpcG2-68 has a big advantage over PcpcG2-172: reducing the volume of the circuit and cheaper to synthesis.
T--XHD-Wuhan-B-China--111(1).png

Please view https://parts.igem.org/Part:BBa_K3921000 for more details.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]