Difference between revisions of "Team:SJTU-BioX-Shanghai/Composite Part"

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                         <div class="my-title h5-my-responsive">总体介绍</div>
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                         <div class="my-title h5-my-responsive">Introduction</div>
                         <p>我们很高兴能为大家呈现这个暑假我们构架的所有关于多因子可视化检测的整合模块</p>
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                         <p>We are so glad to present all the composite parts we have built relevant to our multifactorial visualized detection system during this summer. </p>
                         <p class="my-title2">我们的多因子可视化检测系统是怎么工作的? </p>
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                         <p class="my-title2">How our multifactorial visualized detection system work? </p>
 
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                             <img src="https://static.igem.org/mediawiki/2017/a/a3/T--SJTU-BioX-Shanghai--17210.png" class="img-fluid">
 
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                         <p>大量的色素蛋白可以添加到Target的下游,在这里我们使用amilGFP, amilCP, eforRed, cjBlue这四种色素蛋白。为了响应外界环境的变化,我们需要选择适当的启动子,添加到Target的前面。当想要检测的因子不存在时,Antisense不会被表达,Target中的发卡结构也就不会被打开,下游的色素蛋白就不会表达。当外界因子存在时,响应的启动子就会被激活,Antisense转录进而打开Target中的发卡结构,下游的色素蛋白基因得以表达,颜色显现。</p>
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                         <p>A large variety of chromoproteins can be added on the downstream of Target 1 or Target 3. Here we used amilGFP, amilCP, eforRed, cjBlue four chromoproteins in our detection system. To response to change of outer environment, what
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                            we need to do is just choosing the right promoter on the upstream of Antisense 1 or 3. In the absence of factors of interest, Antisense won’t be transcribed, thus Target hairpin structure won’t be disrupted and downstream chromoprotein
 +
                            won’t be expressed. In the presence of factors, promoter is activated, leading transcription of Antisense and subsequently Target is disrupted, colors of chromoproteins exhibit.</p>
 
                        
 
                        
                         <p>两个基因回路被构建在同一个质粒上,我们使用双表达质粒pCDF-Duet1和pET-Duet1。常表达启动子J23119和Target是提交到公司一起合成的,Antisense和它下游的终止子t500也一样。因为我们的系统有许多优点,所以我们有信心这个系统作为荧光蛋白的替代品会被广泛使用。</p>
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                         <p>The two circuits are constructed into the same plasmid pET-Duet1 or pCDF-Duet1 dual-expression vector. Constitutive promoter J23119 and Target are together synthesized as a whole part and the same as Antisense followed by t500.
                         <p class="my-title2">主要优点:</p>
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                            We have confidence that this system can be widely used as alternative to fluorescence proteins for several merits.</p>
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                         <p class="my-title2">Key advantages:</p>
 
                         <ol type="1" start="1">
 
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                             <li> STAR1和STAR3系统具有很强的正交性,彼此互不影响;</li>
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                             <li> So far, two environmental factors can be detected by our system via STAR 1 and STAR 3 and the two STAR systems won’t affect each other. Their DNA sequences are completely different and Blast result showed there is no significant
                             <li>色素蛋白可被肉眼看到,不需要借助如紫外灯等额外设备;</li>
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                                similarity.</li>
                             <li>我们创造了与之搭配的手机支架和颜色检测APP以更加方便根据颜色得到因子浓度;</li>
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                             <li>Chromoproteins are excellent indicators because they can be seen just by naked eyes, which means no extra equipment, like UV light, is needed.</li>
                             <li>更多关于STAR的好处已经在帝国理工项目中作了讨论(<a target="_blank" href="https://2016.igem.org/Team:Imperial_College/Basic_Part">更多信息</a>.</li>
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                             <li>We have also built a detection platform including a scaffold to place a smartphone and APP to analyze colors that chromoproteins exhibit according to the concentration of factors.</li>
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                             <li>More advantages about STAR itself have been discussed by iGEM16_Imperial and for more information about STAR you can refer to <a target="_blank" href="https://2016.igem.org/Team:Imperial_College/Basic_Part">here</a>.</li>
 
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                         <div class="my-title h5-my-responsive">Parts汇总表</div>
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                         <div class="my-title h5-my-responsive">Parts Table</div>
 
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Revision as of 12:34, 9 December 2017

Composite Parts
Introduction

We are so glad to present all the composite parts we have built relevant to our multifactorial visualized detection system during this summer.

How our multifactorial visualized detection system work?

Figure 1. The STAR-chromoprotein construct.

A large variety of chromoproteins can be added on the downstream of Target 1 or Target 3. Here we used amilGFP, amilCP, eforRed, cjBlue four chromoproteins in our detection system. To response to change of outer environment, what we need to do is just choosing the right promoter on the upstream of Antisense 1 or 3. In the absence of factors of interest, Antisense won’t be transcribed, thus Target hairpin structure won’t be disrupted and downstream chromoprotein won’t be expressed. In the presence of factors, promoter is activated, leading transcription of Antisense and subsequently Target is disrupted, colors of chromoproteins exhibit.

The two circuits are constructed into the same plasmid pET-Duet1 or pCDF-Duet1 dual-expression vector. Constitutive promoter J23119 and Target are together synthesized as a whole part and the same as Antisense followed by t500. We have confidence that this system can be widely used as alternative to fluorescence proteins for several merits.

Key advantages:

  1. So far, two environmental factors can be detected by our system via STAR 1 and STAR 3 and the two STAR systems won’t affect each other. Their DNA sequences are completely different and Blast result showed there is no significant similarity.
  2. Chromoproteins are excellent indicators because they can be seen just by naked eyes, which means no extra equipment, like UV light, is needed.
  3. We have also built a detection platform including a scaffold to place a smartphone and APP to analyze colors that chromoproteins exhibit according to the concentration of factors.
  4. More advantages about STAR itself have been discussed by iGEM16_Imperial and for more information about STAR you can refer to here.
Parts Table
<groupparts>iGEM2017 SJTU-BioX-Shanghai</groupparts>
Reference
  1. Chappell J, Takahashi MK, Lucks JB. 2015. Creating small transcription activating RNAs. Nat Chem Biol 11:214–220.
  2. Meyer, S., Chappell, J., Sankar, S., Chew, R., and Lucks, J. B. (2016) Improving fold activation of small transcription activating RNAs (STARs) with rational RNA engineering strategies Biotechnol. Bioeng. 113, 216.