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| <div class="content_conts"> | | <div class="content_conts"> |
| <!------------------------Liver function and protection------------------------------ --> | | <!------------------------Liver function and protection------------------------------ --> |
− | <h3 class="content_subtitle">Liver function and protection</h3> | + | <h6 style=" margin-top: 15px; |
− | <p class="content_context" style="text-indent:2em; text-align:justify">
| + | margin-left: 25px; |
− | <a href="http://www.discoverymedicine.com/category/species-and-cell-types/bacterium/lactococcus-lactis/">Lactococcus lactis</a>
| + | font-size: 18px; |
− | is one of the most common organisms used in bacterial drug delivery system.
| + | font-family: 'Open Sans', sans-serif;">Design |
− | <a href="http://www.discoverymedicine.com/category/species-and-cell-types/bacterium/lactococcus-lactis/">Lactococcus lactis</a>
| + | and integration of three different modules (gshF, met K and |
− | is a food-grade bacterium that is widely used in the dairy industry. It is a
| + | cwaA)</h6> |
− | Gram-positive bacterium and therefore lacks endotoxic lipopolysaccharides (LPS) which are
| + | <p><img style="width: 70%; margin-top: 3em" src="https://static.igem.org/mediawiki/2018/4/41/T--H14Z1_Hangzhou--project_design_fig1.png"></p> |
− | associated with commonly used
| + | |
− | <a href="http://www.discoverymedicine.com/category/species-and-cell-types/bacterium/gram-negative-bacteria/">Gram-negative bacteria</a>
| + | |
− | . It is also non-invasive and non-commensal | + | |
− | and thus has less potential to trigger immunotolerance or side effects upon prolonged use. The
| + | |
− | bacterium has an established safety profile through its long use in fermented dairy products
| + | |
− | and is considered as a GRAS (generally recognized as safe) microorganism. During the last two
| + | |
− | decades, significant advances have been made in the field of lactococcal genetics and protein
| + | |
− | expression systems. The most commonly used system is the nisin-controlled gene expression | + | |
− | (NICE) system, containing the nisin promoter. Consequently, we have seen the emergence of new
| + | |
− | areas for the application of engineered L. lactis for protein expression,
| + | |
− | <a href="http://www.discoverymedicine.com/category/research-technology/nucleic-acid/gene-delivery/">gene delivery</a>,
| + | |
− | vaccine delivery, and therapeutic
| + | |
− | <a href="http://www.discoverymedicine.com/category/research-technology/drug-delivery-research-technology/">drug delivery.</a>
| + | |
− | </p>
| + | |
| | | |
− | <p class="content_context" style="text-indent:2em; text-align:justify"> | + | <h6 style=" margin-top: 15px; |
− | In the present project, since the oral table administration of GSH and SAM have some
| + | margin-left: 25px; |
− | disadvantages, such as low stability and short life span, here we tried to develop a novel
| + | font-size: 18px; |
− | in-vivo strategy of produce and deliver them simultaneously by using NICE system. In the
| + | font-family: 'Open Sans', sans-serif;">Functional characterization of pNZ-SGC in <i>L.lactis</i> NZ9000</h6> |
− | experiment, two-functional GSH synthetase gene (gshF) and SAM synthetase gene (metK) were in
| + | <p><img style="width: 70%; margin-top: 3em" src="https://static.igem.org/mediawiki/2018/e/e8/T--H14Z1_Hangzhou--project_design_fig2.png"></p> |
− | tandem inserted into the expression vector (pNZ8148), and the resulted plasmid (pNZ8148-SG) was
| + | |
− | employed to construct the target vector pNZ8148-SGC by introducing adhesion factor gene (cwaA).
| + | |
− | This target vector was transformed to get recombinant Lactococcus lacti, which was employed to
| + | |
− | produce our “smart yogurt”.
| + | |
− | </p> | + | |
− | <h3 class="content_subtitle">
| + | |
− | The project is outlined with two stages:
| + | |
− | </h3>
| + | |
− | <!-- ------------------------Stage A---------------------------- -->
| + | |
− | <p class="mt8 content_context"> | + | |
− | <span style="width:120px; font: bold; font-size: 25px">Stage A</span><span>Construction of Lactococcus lactis NZ9000/pNZ-SGC</span>
| + | |
− | </p>
| + | |
− | <p><img style="width: 50%; margin-top: 1em" src="https://static.igem.org/mediawiki/2018/b/be/T--H14Z1_Hangzhou--description_projectOverview_fig1.png"></p>
| + | |
− | <p><img style="width: 50%; margin-top: 1em" src="https://static.igem.org/mediawiki/2018/c/cd/T--H14Z1_Hangzhou--description_projectOverview_fig12.png"></p>
| + | |
− | <p><img style="width: 50%; margin-top: 1em" src="https://static.igem.org/mediawiki/2018/8/87/T--H14Z1_Hangzhou--description_projectOverview_fig3.png"></p> | + | |
| | | |
− | <!-- ------------------------Stage B---------------------------- --> | + | <h6 style=" margin-top: 15px; |
− | <p class="mt8 content_context"> | + | margin-left: 25px; |
− | <span style="width:120px; font: bold; font-size: 25px">Stage B</span><span>Preparation of smart yogurt with our constructed Lactococcus lactis</span> | + | font-size: 18px; |
| + | font-family: 'Open Sans', sans-serif;">Production of “smart yogurt” with our constructed <i>L.lactis</i> |
| + | NZ9000/pNZ-GMcA</h6> |
| + | <p><img style="width: 70%; margin-top: 3em" src="https://static.igem.org/mediawiki/2018/6/65/T--H14Z1_Hangzhou--project_design_fig3.png"></p> |
| + | <p class="content_context" style="text-align:center; font-size:18px"> |
| + | <a style="text-decoration: none; font-size: 18px" href="https://www.nyas.org/ebriefings/cytokine-therapies/">https://www.nyas.org/ebriefings/cytokine-therapies/</a> |
| </p> | | </p> |
− | <p><img style="width: 50%; margin-top: 1em" src="https://static.igem.org/mediawiki/2018/6/66/T--H14Z1_Hangzhou--description_projectOverview_fig4.png"></p>
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| </div> | | </div> |
| </div> | | </div> |