Difference between revisions of "Team:CHINA CD UESTC/Method"
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<h3>Q1: How to get target gene?</h3> | <h3>Q1: How to get target gene?</h3> | ||
− | <p>Four operons related to magnetosome synthesis (<i>mamAB</i>/<i>mamGFDC</i>/<i>mamXY</i>/<i>mms6</i>) and <i>mamW</i>: use the magnetotactic bacteria’s (Magnetospirillum gryphiswaldense MSR -1) genome as template to amplify( | + | <p>Four operons related to magnetosome synthesis (<i>mamAB</i>/<i>mamGFDC</i>/<i>mamXY</i>/<i>mms6</i>) and <i>mamW</i>: use the magnetotactic bacteria’s (Magnetospirillum gryphiswaldense MSR -1) genome as template to amplify(Figure 1). |
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− | <i> | + | <i>laccase</i> gene and <i>RFP</i>: use the DNA fragments from 2015 Kit Plate2 provided by iGEM (Name: <a href="http://parts.igem.org/Part:BBa_K863005">BBa_K863005</a>, <a href="http://parts.igem.org/Part:BBa_E1010">BBa_E1010</a>) and amplify them through common PCR. |
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<h3>Q2: Where are the backbone vectors from?</h3> | <h3>Q2: Where are the backbone vectors from?</h3> | ||
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− | All backbone vectors are purchased from Biotech Corp. They are | + | All backbone vectors are purchased from Biotech Corp. They are pET-28a(Figure 2), pCDFDuet-1(Figure 3) and pACYCDuet-1(Figure 4), the first two aims to carry gene clusters that realize magnetosome generating and the last one is for putting the genes (<i>mamW</i> + <i>RFP</i> + <i>laccase</i>) together. |
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<img src="https://static.igem.org/mediawiki/2015/b/b3/CHINA_CD_UESTC_METHOD02.png" width="60%"> | <img src="https://static.igem.org/mediawiki/2015/b/b3/CHINA_CD_UESTC_METHOD02.png" width="60%"> | ||
<p id="pic_illustration"> | <p id="pic_illustration"> | ||
− | <strong>Figure 2.</strong> This vector backbone ( | + | <strong>Figure 2.</strong> This vector backbone (pET-28a) will be inserted <i>mamAB</i></p> |
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<div class="project_pic"> | <div class="project_pic"> | ||
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− | We obtained the linker information through searching in the Registry of Standard Biological Parts, finally we used two kinds of linkers. | + | We obtained the linker information through searching in the Registry of Standard Biological Parts, finally we used two kinds of linkers(Figure 5). |
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Latest revision as of 02:52, 18 September 2015
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METHOD
We present fundamental details on various methods such as vector design, domain linker selection and choose of restriction enzyme sites used in the experiment on this page. Any questions or advice are welcomed at any time.