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<html>
<link href="http://2017.igem.org/Template:Vilnius-Lithuania/CSSvendor?action=raw&ctype=text/css" rel="stylesheet"/>
<link href="http://2017.igem.org/Template:Vilnius-Lithuania/CSSprod?action=raw&ctype=text/css" rel="stylesheet"/>
</html>


<html>
<div class="wrapper problem-page">
   <div class="container">
     <h1>Present Challenges of<br/> Experimenting With Plasmid Systems</h1>
     <div class="presentation">
       <div class="left">
         <div class="left-steps">
           <div class="left-step is-current" style="display: block">
             <div class="step">1. 1.</div>
             <div class="title">Plasmid Replication </div>
             <p>Origin of replication (Ori) is a sequence where the plasmid replication begins. Plasmids with similar Ori sites use the same replication machinery. </p>
             <h6>Problem #1</h6>
           </div>
           <div class="left-step">
             <div class="step">1. 2.</div>
             <div class="title">Unbalanced Replication</div>
             <p> Replication machinery does not discriminate between similar Ori sites, therefore smaller plasmids are replicated faster and rapidly outgrow larger plasmids, which leads to uneven plasmid ratios in a cell.</p>
             <h6>Problem #1</h6>
           </div>
           <div class="left-step">
             <div class="step">1. 3.</div>
             <div class="title">Plasmid Loss</div>
             <p> This is even more likely to occur when some plasmids are low copy number. When the cell
               divides, low copy plasmids tend to be lost in next generations. </p>
             <h6>Problem #1</h6>
           </div>
           <div class="left-step">
             <div class="step">2. 1.</div>
             <div class="title">Plasmid Incompatibility </div>
             <p>This inability to stably co-maintain different plasmids over a number of generations in the same bacterial cell line is called plasmid incompatibility. One needs to use different types of Ori to avoid this trouble.</p>
             <h6>Problem #2</h6>
           </div>
           <div class="left-step">
             <div class="step">2. 2.</div>
             <div class="title">Plasmid Incompatibility</div>
             <p>However, it often causes trouble due to limited Ori selection options or the lack of good combination of Ori and plasmid's copy number. Imagine you require a system consisting of two - low and high copy number  plasmids. Try picking them from the list.</p>
             <h6>Problem #2</h6>
           </div>
           <div class="left-step">
             <div class="step">2. 3.</div>
             <div class="title">Fixed Copy Number</div>
             <p>Currently no handy method of changing the copy number of particular plasmid exists. It must be either low, or medium, or high.</p>
             <h6>Problem #2</h6>
           </div>
           <div class="left-step">
             <div class="step">2. 4.</div>
             <div class="title">Deficient Responsivity</div>
             <p>There is also no mechanism to adjust the copy number when so is required in any specific
               situation, for example: incoming cellular signals or special patterns.</p>
             <h6>Problem #2</h6>
           </div>
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               <img src="http://2017.igem.org/wiki/images/5/52/T--Vilnius-Lithuania--step-2-1-image1.png" width="381" height="185">
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               <div class="success-text" style="display: none">Success!</div>
               <div class="error-text" style="display: none">Error!</div>
               <div class="interactive-plasmid suc-err">
                 <img src="http://2017.igem.org/wiki/images/4/42/T--Vilnius-Lithuania--step-2-2-dna.png" class="dna">
                 <img src="http://2017.igem.org/wiki/images/5/58/T--Vilnius-Lithuania--step-2-2-int1.png" class="int1" width="42"
                   height="42">
                 <img src="http://2017.igem.org/wiki/images/f/f0/T--Vilnius-Lithuania--step-2-2-int2.png" class="int2" width="42"
                   height="42">
                 <img src="http://2017.igem.org/wiki/images/8/8a/T--Vilnius-Lithuania--step-2-2-int3.png" class="int3" width="42"
                   height="42">
                 <img src="http://2017.igem.org/wiki/images/3/35/T--Vilnius-Lithuania--step-2-2-int4.png" class="int4" width="42"
                   height="42">
                 <img src="http://2017.igem.org/wiki/images/4/44/T--Vilnius-Lithuania--step-2-2-int5.png" class="int5" width="42"
                   height="42">
               </div>
               <div class="success-text-long" style="display: none">
                 As you can see from our example, even when one knows what copy number is needed for system, it is hard to find the right combination of compatible replicons and right copy number.
               </div>
               <div class="error-text-long" style="display: none">
                 <div>Plasmids are incompatible, because they use the same replicons.</div>
                 <div class="button">Try again</div>
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               <img class="i23center" src="http://2017.igem.org/wiki/images/3/3a/T--Vilnius-Lithuania--step-2-3-centermedium.png" width="381" height="185">
               <img class="i23center" src="http://2017.igem.org/wiki/images/6/61/T--Vilnius-Lithuania--step-2-3-centerlow.png" width="381" height="185">
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                   <img src="http://2017.igem.org/wiki/images/2/2d/T--Vilnius-Lithuania--step-2-4-wave.png" width="94" height="63">
                 </div>
                 <img src="http://2017.igem.org/wiki/images/6/66/T--Vilnius-Lithuania--step-2-4-plasmid1.png" class="plasmid" width="48" height="48">
                 <img src="http://2017.igem.org/wiki/images/a/a6/T--Vilnius-Lithuania--step-2-4-plasmid2.png" class="plasmid" width="48" height="48">
                 <img src="http://2017.igem.org/wiki/images/3/39/T--Vilnius-Lithuania--step-2-4-plasmid3.png" class="plasmid" width="50" height="48">
                 <img src="http://2017.igem.org/wiki/images/6/66/T--Vilnius-Lithuania--step-2-4-plasmid1.png" class="plasmid" width="48" height="48">
                 <img src="http://2017.igem.org/wiki/images/a/a6/T--Vilnius-Lithuania--step-2-4-plasmid2.png" class="plasmid" width="48" height="48">
                 <img src="http://2017.igem.org/wiki/images/3/39/T--Vilnius-Lithuania--step-2-4-plasmid3.png" class="plasmid" width="50" height="48">
                 <img src="http://2017.igem.org/wiki/images/6/66/T--Vilnius-Lithuania--step-2-4-plasmid1.png" class="plasmid" width="48" height="48">
                 <img src="http://2017.igem.org/wiki/images/a/a6/T--Vilnius-Lithuania--step-2-4-plasmid2.png" class="plasmid" width="48" height="48">
                 <img src="http://2017.igem.org/wiki/images/3/39/T--Vilnius-Lithuania--step-2-4-plasmid3.png" class="plasmid" width="50" height="48">
                 <img src="http://2017.igem.org/wiki/images/6/66/T--Vilnius-Lithuania--step-2-4-plasmid1.png" class="plasmid" width="48" height="48">
                 <img src="http://2017.igem.org/wiki/images/4/42/T--Vilnius-Lithuania--step-2-2-dna.png" class="dna">
               </div>
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               <h2>Did this look smart, easy or convenient?</h2>
               <p>We think it is time for a change.</p>
             </div>
           </div>
         </div>
         <div class="mob-prev" style="background-image: url('http://2017.igem.org/wiki/images/a/a0/T--Vilnius-Lithuania--step-1-1-arrow.png')"></div>
         <div class="mob-next" style="background-image: url('http://2017.igem.org/wiki/images/a/a0/T--Vilnius-Lithuania--step-1-1-arrow.png')"></div>
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       <div class="right">
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                   <img src="http://2017.igem.org/wiki/images/f/fa/T--Vilnius-Lithuania--step-1-1-boxline1.png" width="42"
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                 <div class="box-line-name">Plasmid A</div>
               </div>
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                   <img src="http://2017.igem.org/wiki/images/1/1a/T--Vilnius-Lithuania--step-1-1-boxline2.png" width="42"
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                 </div>
                 <div class="box-line-name">Plasmid B</div>
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                   <img src="http://2017.igem.org/wiki/images/f/f9/T--Vilnius-Lithuania--step-1-1-boxline3.png" width="12"
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                 </div>
                 <div class="box-line-name">Origin of replication</div>
               </div>
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                   <img src="http://2017.igem.org/wiki/images/3/3d/T--Vilnius-Lithuania--step-1-1-boxline4.png" width="19"
                     height="19">
                 </div>
                 <div class="box-line-name">Replication machinery</div>
               </div>
             </div>
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               <div class="box-line">
                 <div class="imgcont">
                   <img src="http://2017.igem.org/wiki/images/f/fa/T--Vilnius-Lithuania--step-1-1-boxline1.png" width="42"
                     height="42">
                 </div>
                 <div class="box-line-name">Plasmid A</div>
               </div>
               <div class="box-line">
                 <div class="imgcont">
                   <img src="http://2017.igem.org/wiki/images/1/1a/T--Vilnius-Lithuania--step-1-1-boxline2.png" width="42"
                     height="42">
                 </div>
                 <div class="box-line-name">Plasmid B</div>
               </div>
               <div class="box-line">
                 <div class="imgcont">

2022年11月20日 (日) 09:13的版本

取自“https://hmoegirl.com/index.php?title=Widget:新样式&oldid=63008”