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

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       <h2>Type IIs Restriction Endonuclease</h2>
 
       <h2>Type IIs Restriction Endonuclease</h2>
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<p> 限制性内切酶可以根据其酶切方式的不同分为4类，分别为TypeI、TypeII、TypeIII和TypeIV。我们常用的限制性核酸内切酶，如EcoRI，都属于II型限制性核酸内切酶，这一类限制性内切酶可以识别特异性的4～8个核苷酸序列，识别序列反向重复，且切割位点位于识别位点内，酶切后产生粘性末端或者平末端，其切割示意图如下
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<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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但是，Type IIS限制性内切酶则是一种相对特殊的酶，与普通的Type II限制性内切酶相比，这类酶会在识别位点下游切割DNA，从酶的三维结构分析，这是由于Type IIS限制性内切酶的识别区域和催化区域被一个多肽连接子分开。
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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>
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TypeIIS限制性内切酶的优点在于其对于切割序列没有要求，识别序列外的位置可以是任意序列，多个DNA片段可以通过互补的末端进行组装，这种组装方法又叫做Goldengate。
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</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>
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Revision as of 04:02, 30 September 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:

    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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