Difference between revisions of "Team:Jilin China/Construction"

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       <h2>Type IIs Restriction Endonuclease</h2>
 
       <h2>Type IIs Restriction Endonuclease</h2>
 
<p>Restriction enzymes can be divided into four categories according to their different enzyme digestion methods, named Type I, Type II, Type III and Type IV. Our commonly used restriction endonucleases, such as EcoRI, belong to type II restriction endonucleases, which recognize specific 4 to 8 nucleotide sequences . The recognized sequence is inverted repeats and the cleavage site is located in the recognition site. After enzymatic cleavage and the cleavage end or blunt end is produced. The cutting diagram is as follows:</p>
 
<p>Restriction enzymes can be divided into four categories according to their different enzyme digestion methods, named Type I, Type II, Type III and Type IV. Our commonly used restriction endonucleases, such as EcoRI, belong to type II restriction endonucleases, which recognize specific 4 to 8 nucleotide sequences . The recognized sequence is inverted repeats and the cleavage site is located in the recognition site. After enzymatic cleavage and the cleavage end or blunt end is produced. The cutting diagram is as follows:</p>
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<img src="https://static.igem.org/mediawiki/2018/4/48/T--Jilin_China--Team--Members--WangZihao.jpeg" alt="Wang Zihao " />
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      <polyline class="f cut" points="196.12 59.76 196.12 95.54 308.12 95.54 308.12 128.26"/>
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      <path d="M131,150.41l-.82,1.18a2.44,2.44,0,0,0-.9-.48,3.88,3.88,0,0,0-1.27-.23,2.71,2.71,0,0,0-2.17,1,4,4,0,0,0-.8,2.62,3.57,3.57,0,0,0,.82,2.52,3,3,0,0,0,2.27.86,3.75,3.75,0,0,0,2.27-.87l.66,1.41a6,6,0,0,1-3.33.87,4.27,4.27,0,0,1-3.19-1.29,4.84,4.84,0,0,1-1.25-3.5,5,5,0,0,1,1.3-3.61,4.71,4.71,0,0,1,3.57-1.37,4.85,4.85,0,0,1,1.59.31A4.76,4.76,0,0,1,131,150.41Z" transform="translate(0 -2.86)"/>
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      <path d="M149.74,159.06l-3.44-5.52q-.59,0-1.86-.09v5.61h-1.76V146.19l1.37-.07c.85,0,1.46-.07,1.85-.07,3.09,0,4.64,1.24,4.64,3.7a3.49,3.49,0,0,1-.75,2.18,3.09,3.09,0,0,1-1.77,1.21l3.73,5.92Zm-5.3-11.29V152a8.69,8.69,0,0,0,1.23.09,4,4,0,0,0,2.3-.51,2.09,2.09,0,0,0,.72-1.82,1.66,1.66,0,0,0-.77-1.55,4.76,4.76,0,0,0-2.44-.48C145.3,147.68,145,147.71,144.44,147.77Z" transform="translate(0 -2.86)"/>
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    </g>
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</svg>
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</div>
 +
<h1>R IIs</h1>
 +
<div class="R2s_animation">
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<svg xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink" width="100%" viewBox="0 0 505.59 171.73">
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  <defs>
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    <style>
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      .R2s_animation .a,.R2s_animation .g {
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        fill: #29abe2;
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      }
 +
 
 +
      .R2s_animation .a,.R2s_animation .c {
 +
        opacity: 0.51;
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      }
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 +
      .R2s_animation .b,.R2s_animation .c {
 +
        fill: #22b573;
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      }
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      .R2s_animation .d,.R2s_animation .i {
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        fill: none;
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        stroke-linecap: round;
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        stroke-miterlimit: 10;
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      }
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      .R2s_animation .d {
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        stroke-width: 7.45px;
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        stroke: url(#a);
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      }
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      .R2s_animation .e {
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        fill: #9000a5;
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      }
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 +
      .R2s_animation .f {
 +
        fill: #fff;
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      }
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 +
      .R2s_animation .h {
 +
        fill: #ed1c24;
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      }
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 +
      .R2s_animation .i {
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        stroke: #1b1464;
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        stroke-width: 5px;
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        stroke-dasharray: 3 12;
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      }
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.R2s_animation .right{
 +
transition:all 0.3s ease;
 +
}
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.R2s_animation .enzyme{
 +
opacity:0;
 +
transform: translateY(-25%);
 +
transition:all 0.3s ease;
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}
 +
.R2s_animation:hover .right{
 +
transform: translateX(25%);
 +
transition:all 0.3s ease 0.6s;
 +
}
 +
.R2s_animation:hover .enzyme{
 +
opacity: 1;
 +
transform: translateY(0);
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}
 +
.R2s_animation:hover .cut{
 +
opacity: 0;
 +
transition: 0.3s all ease 0.6s;
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}
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      <stop offset="0" stop-color="#29abe2" stop-opacity="0.51"/>
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      <stop offset="0.99" stop-color="#22b573" stop-opacity="0.51"/>
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    </linearGradient>
 +
  </defs>
 +
  <title>R IIs</title>
 +
  <g>
 +
    <g>
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      <path d="M125.7,159.5l.59-1.58a4.51,4.51,0,0,0,2.25.91c1,0,1.55-.43,1.55-1.3,0-.62-.5-1.15-1.5-1.6a11.08,11.08,0,0,1-1.55-.8,2.94,2.94,0,0,1-.69-.62,2.23,2.23,0,0,1-.44-.73,2.32,2.32,0,0,1-.15-.84,2.13,2.13,0,0,1,.85-1.81,3.45,3.45,0,0,1,2.2-.65,6.92,6.92,0,0,1,2.59.65l-.47,1.54a3.24,3.24,0,0,0-2-.79,1.78,1.78,0,0,0-1,.28.86.86,0,0,0-.41.72,1.48,1.48,0,0,0,1,1.37l1.19.54a4.25,4.25,0,0,1,1.6,1.14,2.52,2.52,0,0,1,.5,1.61,2.38,2.38,0,0,1-.88,2,3.83,3.83,0,0,1-2.46.72A5.64,5.64,0,0,1,125.7,159.5Z" transform="translate(0 -2.9)"/>
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      <path d="M148.92,160.06V147.19h1.76v12.87Z" transform="translate(0 -2.9)"/>
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    <g class="enzyme">
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      <g>
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        <path class="b" d="M235.91,18.41l-4.1-6.59c-.47,0-1.21,0-2.22-.11v6.7h-2.1V3.05L229.12,3c1-.06,1.75-.08,2.21-.08q5.53,0,5.53,4.4A4.18,4.18,0,0,1,236,9.9a3.61,3.61,0,0,1-2.12,1.44l4.46,7.07ZM229.59,4.94v5a10.61,10.61,0,0,0,1.47.11,4.87,4.87,0,0,0,2.74-.61,2.5,2.5,0,0,0,.86-2.17,2,2,0,0,0-.92-1.86,5.88,5.88,0,0,0-2.92-.56C230.61,4.84,230.2,4.87,229.59,4.94Z" transform="translate(0 -2.9)"/>
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        <path class="b" d="M246.71,18.41V3.05h2.1V18.41Z" transform="translate(0 -2.9)"/>
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        <path class="b" d="M252.69,18.41V3.05h2.09V18.41Z" transform="translate(0 -2.9)"/>
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        <path class="b" d="M257.4,17.74l.71-1.89a5.36,5.36,0,0,0,2.68,1.09c1.23,0,1.84-.52,1.84-1.55,0-.74-.59-1.38-1.78-1.91a15,15,0,0,1-1.85-1,4.22,4.22,0,0,1-.83-.74,3.19,3.19,0,0,1-.52-.88,3,3,0,0,1-.17-1,2.58,2.58,0,0,1,1-2.16A4.18,4.18,0,0,1,261.11,7a8.47,8.47,0,0,1,3.1.78l-.57,1.84a3.86,3.86,0,0,0-2.38-.94,2,2,0,0,0-1.2.34,1,1,0,0,0-.49.85,1.76,1.76,0,0,0,1.23,1.63l1.43.65a5,5,0,0,1,1.9,1.36,3,3,0,0,1,.6,1.92,2.83,2.83,0,0,1-1.06,2.36,4.53,4.53,0,0,1-2.93.86A6.7,6.7,0,0,1,257.4,17.74Z" transform="translate(0 -2.9)"/>
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    </g>
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    <g class="right">
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      <g>
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<!--预警结束[Warning End]-->
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<p>However, compared with the common Type II restriction enzymes the Type IIS restriction endonuclease is a relatively specific enzyme that cleaves DNA downstream of the recognition site, from the three-dimensional structure analysis of the enzyme. This is due to the fact that the recognition and catalytic regions of the Type IIS restriction enzyme are separated by a polypeptide linker.</p>
 
<p>However, compared with the common Type II restriction enzymes the Type IIS restriction endonuclease is a relatively specific enzyme that cleaves DNA downstream of the recognition site, from the three-dimensional structure analysis of the enzyme. This is due to the fact that the recognition and catalytic regions of the Type IIS restriction enzyme are separated by a polypeptide linker.</p>
 
<p> The advantage of the TypeII S restriction enzyme is that it has no requirement to a cleavage sequence. The cutting position can be any sequence, and multiple DNA fragments can be assembled through complementary ends. This assembly method is also called GoldenGate Assembly.</p>
 
<p> The advantage of the TypeII S restriction enzyme is that it has no requirement to a cleavage sequence. The cutting position can be any sequence, and multiple DNA fragments can be assembled through complementary ends. This assembly method is also called GoldenGate Assembly.</p>

Revision as of 12:32, 9 October 2018

CONSTRUCTION


Construction

  • Type IIs Restriction Endonuclease

    Restriction enzymes can be divided into four categories according to their different enzyme digestion methods, named Type I, Type II, Type III and Type IV. Our commonly used restriction endonucleases, such as EcoRI, belong to type II restriction endonucleases, which recognize specific 4 to 8 nucleotide sequences . The recognized sequence is inverted repeats and the cleavage site is located in the recognition site. After enzymatic cleavage and the cleavage end or blunt end is produced. The cutting diagram is as follows:

    R II

    R IIs

    R IIs

    However, compared with the common Type II restriction enzymes the Type IIS restriction endonuclease is a relatively specific enzyme that cleaves DNA downstream of the recognition site, from the three-dimensional structure analysis of the enzyme. This is due to the fact that the recognition and catalytic regions of the Type IIS restriction enzyme are separated by a polypeptide linker.

    The advantage of the TypeII S restriction enzyme is that it has no requirement to a cleavage sequence. The cutting position can be any sequence, and multiple DNA fragments can be assembled through complementary ends. This assembly method is also called GoldenGate Assembly.

  • References

    • Dai H, Wang Y, Lu X, et al. Chimeric antigen receptors modified T-cells for cancer therapy[J]. JNCI: Journal of the National Cancer Institute, 2016, 108(7).
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