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<p> </p>
<p>(3) pTrc-Spe1-mETP-Xho1-APEX2 protein his column purification</p>
<p>(E.Coli expression vector)</p>
<p>[[File:Ex13.jpg|400px]]</p>
<p> </p>
<p>His column purification Comassie blue staining</p>
<p>[[File:Ex14.png|400px]]</p>
<p>(3) pTrc-Spe1-mETP-Xho1-APEX2 protein</p>
<p> </p>
<p>#19 size exclusion(purification)</p>
<p>[[File:Ex15.jpg|400px]]</p>
<p>(3) pTrc-Spe1-mETP-Xho1-APEX2 protein</p>
<p> </p>
<p>(1)pRA-GFP-pRL-mETP-SpyTag-Stop-Xho1 (mammalian expression vector)</p>
<p>& (2) pTrc-Spe1-SpyCatcher-Xho1-APEX2</p>
<p>(E.Coli expression vector) </p>
<p>Binding test</p>
<p>(protein to protein)</p>
<p>[[File:Ex16.png|400px]] [[File:Ex17.jpg|400px]] </p>
<p>* pRA-mETP-SpyTag_CM 10ul(concentrated)</p>
<p>pTrc-Spycatcher 200ng, 500ng, 100ng, 200ng, 500ng, 1000ng</p>
<p> binding test(Anti His) (15%)</p>
<p>* pRA-mETP-SpyTag_CM 10ul(concentrated)</p>
<p>pTrc-Spycatcher 200ng, 500ng, 100ng, 200ng, 500ng, 1000ng</p>
<p>binding test(Anti mETP(Tx621))(15%)</p>
<p> </p>
<p>Further study & Prospect</p>
<p> </p>
<p>-Do protein interaction experiment in cell to see whether it is well connected in vitro</p>
<p>-</p>
<p>-</p>
<p>-Find the protein that is correlated with mETP through ‘mass spectroscopy’ technique</p>
<p> </p>
<p> -> Therefore, we can identify unknown ‘protein’ and its gene and their location.</p>
<p> </p>
<p>-Applied to Genomics</p>
<p> </p>
<p> -> Analyze how different, how overexpressed, how different of unknown ‘protein’ gene between obese people to lean people by genome sequencing of certain region of unknown ‘protein’ gene</p>
<p> </p>
<p> </p>
<p>Reference</p>
<p> </p>
<p>- Definition of Proteomics <a href="http://biolecture.org/index.php/Proteomics">http://</a><a href="http://biolecture.org/index.php/Proteomics">biolecture.org/index.php/Proteomics</a></p>
<p>- Definition of Omics <a href="http://biolecture.org/index.php/Omics">http</a><a href="http://biolecture.org/index.php/Omics">://</a><a href="http://biolecture.org/index.php/Omics">biolecture.org/index.php/Omics</a></p>
<p>-Bhak, J. (2016, 6 13). Openfree biolecture. Retrieved from Openfree biolecture: <u><a href="http://biolecture.org/index.php/SELF:_Self_evaluating_learning_framework">http://biolecture.org/index.php/SELF:_Self_evaluating_learning_framework</a></u></p>
<p>-Endotrophin triggers adipose tissue fibrosis and metabolic dysfunction. <u><a href="https://www.ncbi.nlm.nih.gov/pubmed/24647224">https://www.ncbi.nlm.nih.gov/pubmed/24647224</a></u></p>
<p>-Adipocyte-derived endotrophin promotes malignant tumor progression</p>
<p><u><a href="https://www.jci.org/articles/view/63930">https://www.jci.org/articles/view/63930</a></u></p>
<p>-Secrets of a covalent interaction for biomaterials and biotechnology: SpyTag and SpyCatcher.</p>
<p><u><a href="https://www.ncbi.nlm.nih.gov/pubmed/26517567">https://www.ncbi.nlm.nih.gov/pubmed/26517567</a></u></p>
<p>-Directed evolution of APEX2 for electron microscopy and proximity labeling.</p>
<p><u><a href="https://www.ncbi.nlm.nih.gov/pubmed/25419960">https://www.ncbi.nlm.nih.gov/pubmed/25419960</a></u></p>
<p>-From genomics to proteomics (Nature Review)</p>
<p><u><a href="http://www.nature.com/nature/journal/v422/n6928/full/nature01510.html">http://www.nature.com/nature/journal/v422/n6928/full/nature01510.html</a></u></p>
<p>-Innovation: Metabolomics: the apogee of the omics trilogy</p>
<p><u><a href="http://www.nature.com/nrm/journal/v13/n4/abs/nrm3314.html">http://www.nature.com/nrm/journal/v13/n4/abs/nrm3314.html</a></u></p>
<p>-Mass-spectrometric exploration of proteome structure and function</p>
<p><u><a href="http://www.nature.com/nature/journal/v537/n7620/abs/nature19949.html">http://www.nature.com/nature/journal/v537/n7620/abs/nature19949.html</a></u></p>
<p> </p>