Difference between revisions of "Team:TecCEM/SelectionMarkerParts"
| Line 891: | Line 891: | ||
| − | + | <main id="PAGES_CONTAINER" tabindex="-1"><div id="SITE_PAGES"><div class="_2YGAo"><div id="tv0y0" class="_1FOTJ _3LL-w"><div class="_3jgRX"></div><div class="_2ufc7"><div id="Containertv0y0" class="_1KV2M"><div data-mesh-id="Containertv0y0inlineContent" data-testid="inline-content" class=""><div data-mesh-id="Containertv0y0inlineContent-gridContainer" data-testid="mesh-container-content"><div id="comp-kuyuk58h" class="_1Q9if" data-testid="richTextElement"><h1 class="font_0" style="font-size:60px"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold">Selection Marker Parts</span></h1></div><div id="comp-kuyuk58n" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:40px"><span style="font-family:wfont_093ca4_4dcfda9e6d1242458c6bd410b0b78ced,wf_4dcfda9e6d1242458c6bd410b,orig_nunito_regular">Biobricks to revolutionize the Safety in the Lab</span></h2></div><div id="comp-kuyuk58q" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:20px; text-align:justify"><span class="color_14"><span style="font-size:20px"><span style="font-family:wfont_2ce676_53d594d91a1845238779d39b6e80ae45,wf_53d594d91a1845238779d39b6,orig_nunito_extralight">The design of these parts was made thinking about a novel selection marker and a modified strain of <span style="font-style:italic;">E. coli </span>we will use to substitute the use of antibiotics in genetic engineering protocols. The idea is to use this parts to generate a knockout strain from <span style="font-style:italic;">E. coli</span> DH5⍺ genes oxyR, gldA and glpK and substitute with aldO, mRFP and PuDHT genes. We also designed a part that forms a plasmid and acts as a selection cassette. AldO will convert glycerol to peroxide and D-glyceric acid, katE will convert peroxide to water and oxygen and PuDHT will convert D-glyceric acid into pyruvate. We will briefly explore the functionality of each part, as well as key aspects of its design.</span></span></span></h2></div><section id="comp-kuyuk591" class="_3d64y"><div id="bgLayers_comp-kuyuk591" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuyuk591" class="_2GUhU"></div></div><div data-testid="columns" class="_1uldx"><div id="comp-kuyuk5951" class="_1vNJf"><div id="bgLayers_comp-kuyuk5951" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuyuk5951" class="_2GUhU"></div></div><div data-mesh-id="comp-kuyuk5951inlineContent" data-testid="inline-content" class=""><div data-mesh-id="comp-kuyuk5951inlineContent-gridContainer" data-testid="mesh-container-content"><div id="comp-kuyuk59a1" class="_1Q9if" data-testid="richTextElement"><p class="font_8" style="font-size: 31px; text-align: center;"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold"><span style="font-size:31px"><span style="font-weight:bold">BBa_K3809001: Novel Selection Cassette katE</span></span></span></p></div><div id="comp-kuyuqcju" class="XUUsC" title=""><div data-testid="linkElement" class="xQ_iF"><wix-image id="img_comp-kuyuqcju" class="_1-6YJ _1Fe8-" data-image-info="{"containerId":"comp-kuyuqcju","displayMode":"fill","imageData":{"width":624,"height":83,"uri":"d211b2_42a61dc42fcf428fb49cec342af04384~mv2.png","name":"T--TecCEM--PartsFig5.png","displayMode":"fill"}}" data-bg-effect-name="" data-is-svg="false" data-is-svg-mask="false" data-image-zoomed="" data-has-ssr-src="" data-src="https://static.wixstatic.com/media/d211b2_42a61dc42fcf428fb49cec342af04384~mv2.png/v1/fill/w_790,h_105,al_c,lg_1,q_85/T--TecCEM--PartsFig5.webp"><img src="https://static.igem.org/mediawiki/2021/d/db/T--TecCEM--PartsFig5.png" alt="T--TecCEM--PartsFig5.png" style="width:632px;height:84px;object-fit:cover;object-position:50% 50%"></wix-image></div></div><div id="comp-kuyunepk" class="_1Q9if" data-testid="richTextElement"><p class="font_8" style="font-size: 18px; text-align: center;"><span style="font-size:18px;"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold"><span style="font-weight:bold">Figure 1.</span></span></span></p></div><div id="comp-kuyuk59c" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:17px; text-align:center"><span style="font-size:17px"><span class="color_15"><span style="font-family:wfont_069f35_23de2075afcc42cc848bac4f05c000f8,wf_23de2075afcc42cc848bac4f0,orig_roboto">Map of <span style="text-decoration:underline;"><a href="http://parts.igem.org/Part:BBa_K3809001" target="_blank" rel="noreferrer noopener">BBa_K3809001</a></span>. Created with </span></span><a href="https://www.snapgene.com/" target="_blank" rel="noreferrer noopener"><span style="color:#001489"><span style="text-decoration:underline"><span style="font-family:wfont_069f35_23de2075afcc42cc848bac4f05c000f8,wf_23de2075afcc42cc848bac4f0,orig_roboto">Snapgene.</span></span></span></a></span></h2></div><div id="comp-kuyuoyy1" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:20px; text-align:justify"><span class="color_15"><span style="font-size:20px"><span style="font-family:wfont_2ce676_53d594d91a1845238779d39b6e80ae45,wf_53d594d91a1845238779d39b6,orig_nunito_extralight">This part codes for the E. coli catalase gene katE which converts peroxide into water and oxygen. It is meant to be used with a pUC backbone we designed in which the part acts as a selection marker for the modified strain. For more information, click on the part name.</span></span></span></h2></div></div></div></div></div></section><section id="comp-kuyuk59m" class="_3d64y"><div id="bgLayers_comp-kuyuk59m" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuyuk59m" class="_2GUhU"></div></div><div data-testid="columns" class="_1uldx"><div id="comp-kuyuk59o1" class="_1vNJf"><div id="bgLayers_comp-kuyuk59o1" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuyuk59o1" class="_2GUhU"></div></div><div data-mesh-id="comp-kuyuk59o1inlineContent" data-testid="inline-content" class=""><div data-mesh-id="comp-kuyuk59o1inlineContent-gridContainer" data-testid="mesh-container-content"><div id="comp-kuyuk59q2" class="_1Q9if" data-testid="richTextElement"><h1 class="font_0" style="font-size: 33px; text-align: center;"><span style="color:#FAFAFA"><span style="font-size:33px"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold">BBa_K3809003: katE regulated by BBa_J23100</span></span></span></h1></div><div id="comp-kuyzoya3" class="XUUsC" title=""><div data-testid="linkElement" class="xQ_iF"><wix-image id="img_comp-kuyzoya3" class="_1-6YJ _1Fe8-" data-image-info="{"containerId":"comp-kuyzoya3","displayMode":"fill","imageData":{"width":936,"height":132,"uri":"d211b2_e511daa612014087acb6a3f8092c3ea0~mv2.png","name":"T--TecCEM--figura2.png","displayMode":"fill"}}" data-bg-effect-name="" data-is-svg="false" data-is-svg-mask="false" data-image-zoomed="" data-has-ssr-src="" data-src="https://static.wixstatic.com/media/d211b2_e511daa612014087acb6a3f8092c3ea0~mv2.png/v1/fill/w_793,h_111,al_c,q_85,usm_0.66_1.00_0.01/T--TecCEM--figura2.webp"><img src="https://static.igem.org/mediawiki/2021/3/39/T--TecCEM--figura2.png" alt="T--TecCEM--figura2.png" style="width:634px;height:89px;object-fit:cover;object-position:50% 50%"></wix-image></div></div><div id="comp-kuyz626q" class="_1Q9if" data-testid="richTextElement"><p class="font_8" style="font-size:18px; text-align:center"><span style="color:#FFFFFF"><span style="font-size:18px"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold"><span style="font-weight:bold">Figure 2.</span></span></span></span></p></div><div id="comp-kuyz626z" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:17px; text-align:center"><span style="color:#FFFFFF"><span style="font-size:17px"><span style="font-family:wfont_069f35_23de2075afcc42cc848bac4f05c000f8,wf_23de2075afcc42cc848bac4f0,orig_roboto">Map of <span style="text-decoration:underline;"><a href="http://parts.igem.org/Part:BBa_K3809003" target="_blank" rel="noreferrer noopener">BBa_K3809003.</a></span> Created with <span style="font-weight:bold"><span style="text-decoration:underline"><a href="https://www.snapgene.com/" target="_blank" rel="noreferrer noopener">Snapgene</a></span></span></span></span></span></h2></div><div id="comp-kuyz6275" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:20px; text-align:justify"><span style="color:#FFFFFF"><span style="font-size:20px"><span style="font-family:wfont_2ce676_53d594d91a1845238779d39b6e80ae45,wf_53d594d91a1845238779d39b6,orig_nunito_extralight">This Biobrick contains a katE gene with an RBS (BBa_B0030), spacer (BBa_B0040), terminator (BBa_B0010) and promoter (BBa_J23100). It was designed to be expressed as part of a plasmid, to act as a selection cassette for a modified strain of <span style="font-style:italic;">E. coli.</span></span></span></span></h2></div></div></div></div></div></section><section id="comp-kuyzptum" class="_3d64y"><div id="bgLayers_comp-kuyzptum" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuyzptum" class="_2GUhU"></div></div><div data-testid="columns" class="_1uldx"><div id="comp-kuyzptvw" class="_1vNJf"><div id="bgLayers_comp-kuyzptvw" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuyzptvw" class="_2GUhU"></div></div><div data-mesh-id="comp-kuyzptvwinlineContent" data-testid="inline-content" class=""><div data-mesh-id="comp-kuyzptvwinlineContent-gridContainer" data-testid="mesh-container-content"><div id="comp-kuyzptw02" class="_1Q9if" data-testid="richTextElement"><p class="font_8" style="font-size:31px; text-align:center"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold"><span style="font-size:31px"><span style="font-weight:bold">BBa_K3809004: aldO</span></span></span></p></div><div id="comp-kuyzti60" class="XUUsC" title=""><div data-testid="linkElement" class="xQ_iF"><wix-image id="img_comp-kuyzti60" class="_1-6YJ _1Fe8-" data-image-info="{"containerId":"comp-kuyzti60","displayMode":"fill","imageData":{"width":936,"height":124,"uri":"d211b2_8e2baca562f74b309072e3b8d8900bea~mv2.png","name":"T--TecCEM--fig3.png","displayMode":"fill"}}" data-bg-effect-name="" data-is-svg="false" data-is-svg-mask="false" data-image-zoomed="" data-has-ssr-src="" data-src="https://static.wixstatic.com/media/d211b2_8e2baca562f74b309072e3b8d8900bea~mv2.png/v1/fill/w_786,h_104,al_c,q_85,usm_0.66_1.00_0.01/T--TecCEM--fig3.webp"><img src=" https://static.igem.org/mediawiki/2021/e/e9/T--TecCEM--fig3.png" alt="T--TecCEM--fig3.png" style="width:629px;height:83px;object-fit:cover;object-position:50% 50%"></wix-image></div></div><div id="comp-kuyzptw32" class="_1Q9if" data-testid="richTextElement"><p class="font_8" style="font-size:18px; text-align:center"><span style="font-size:18px"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold"><span style="font-weight:bold">Figure 3.</span></span></span></p></div><div id="comp-kuyzptw5" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:17px; text-align:center"><span style="font-size:17px"><span class="color_15"><span style="font-family:wfont_069f35_23de2075afcc42cc848bac4f05c000f8,wf_23de2075afcc42cc848bac4f0,orig_roboto">Map of <span style="text-decoration:underline;"><a href="http://parts.igem.org/Part:BBa_K3809004" target="_blank" rel="noreferrer noopener">BBa_K3809004</a></span>. Created with <span style="text-decoration:underline"><a href="https://www.snapgene.com/" target="_blank" rel="noreferrer noopener"><span style="font-weight:bold">Snapgene</span>.</a></span></span></span></span></h2></div><div id="comp-kuyzptw6" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:20px; text-align:justify"><span class="color_15"><span style="font-size:20px"><span style="font-family:wfont_2ce676_53d594d91a1845238779d39b6e80ae45,wf_53d594d91a1845238779d39b6,orig_nunito_extralight">This sequence codes for the enzyme alditol oxidase from the organism <span style="font-style:italic">Streptomyces</span> <span style="font-style:italic;">coelicolor.</span> It was designed as part of a novel selection method in which the enzyme converts glycerol into D-glycerate. </span></span></span></h2></div></div></div></div></div></section><section id="comp-kuyzvwo8" class="_3d64y"><div id="bgLayers_comp-kuyzvwo8" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuyzvwo8" class="_2GUhU"></div></div><div data-testid="columns" class="_1uldx"><div id="comp-kuyzvwpb1" class="_1vNJf"><div id="bgLayers_comp-kuyzvwpb1" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuyzvwpb1" class="_2GUhU"></div></div><div data-mesh-id="comp-kuyzvwpb1inlineContent" data-testid="inline-content" class=""><div data-mesh-id="comp-kuyzvwpb1inlineContent-gridContainer" data-testid="mesh-container-content"><div id="comp-kuyzvwpe" class="_1Q9if" data-testid="richTextElement"><h1 class="font_0" style="font-size:33px; text-align:center"><span style="color:#FAFAFA"><span style="font-size:33px"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold">BBa_K3809005: aldO + glpK homology regions</span></span></span></h1></div><div id="comp-kuyzyn6p" class="XUUsC" title=""><div data-testid="linkElement" class="xQ_iF"><wix-image id="img_comp-kuyzyn6p" class="_1-6YJ _1Fe8-" data-image-info="{"containerId":"comp-kuyzyn6p","displayMode":"fill","imageData":{"width":936,"height":128,"uri":"d211b2_0e562adb665443429d88c3ab72d5078c~mv2.png","name":"T--TecCEM--fig4.png","displayMode":"fill"}}" data-bg-effect-name="" data-is-svg="false" data-is-svg-mask="false" data-image-zoomed="" data-has-ssr-src="" data-src="https://static.wixstatic.com/media/d211b2_0e562adb665443429d88c3ab72d5078c~mv2.png/v1/fill/w_750,h_104,al_c,q_85,usm_0.66_1.00_0.01/T--TecCEM--fig4.webp"><img src="https://static.igem.org/mediawiki/2021/8/8a/T--TecCEM--fig4.png" alt="T--TecCEM--fig4.png" style="width:600px;height:83px;object-fit:cover;object-position:50% 50%"></wix-image></div></div><div id="comp-kuyzvwpf1" class="_1Q9if" data-testid="richTextElement"><p class="font_8" style="font-size:18px; text-align:center"><span style="color:#FFFFFF"><span style="font-size:18px"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold"><span style="font-weight:bold">Figure 4.</span></span></span></span></p></div><div id="comp-kuyzvwph" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:17px; text-align:center"><span style="color:#FFFFFF"><span style="font-size:17px"><span style="font-family:wfont_069f35_23de2075afcc42cc848bac4f05c000f8,wf_23de2075afcc42cc848bac4f0,orig_roboto">Map of <span style="text-decoration:underline;"><a href="http://parts.igem.org/Part:BBa_K3809005" target="_blank" rel="noreferrer noopener">BBa_K3809005. </a></span>Created with <span style="text-decoration:underline"><a href="https://www.snapgene.com/" target="_blank" rel="noreferrer noopener"><span style="font-weight:bold">Snapgene</span></a></span></span></span></span></h2></div><div id="comp-kuyzvwpi" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:20px; text-align:justify"><span style="color:#FFFFFF"><span style="font-size:20px"><span style="font-family:wfont_2ce676_53d594d91a1845238779d39b6e80ae45,wf_53d594d91a1845238779d39b6,orig_nunito_extralight">This part corresponds to the coding sequence of alditol oxidase. It is regulated by the BBa_J23100 promoter and has a BBa_B0030 RBS and an rrnB T1 terminator BBa_B0010. It also has a spacer between the promoter and the RBS (BBa_B0040). The sequence contains two homology regions for the gene glycerol kinase (glpK) of E. coli, which is a key enzyme in the glycerol metabolic pathway.</span></span></span></h2></div><section id="comp-kuyzyyra" class="_3d64y"><div id="bgLayers_comp-kuyzyyra" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuyzyyra" class="_2GUhU"></div></div><div data-testid="columns" class="_1uldx"><div id="comp-kuyzyysj" class="_1vNJf"><div id="bgLayers_comp-kuyzyysj" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuyzyysj" class="_2GUhU"></div></div><div data-mesh-id="comp-kuyzyysjinlineContent" data-testid="inline-content" class=""><div data-mesh-id="comp-kuyzyysjinlineContent-gridContainer" data-testid="mesh-container-content"><div id="comp-kuyzyysl2" class="_1Q9if" data-testid="richTextElement"><p class="font_8" style="font-size:31px; text-align:center"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold"><span style="font-size:31px"><span style="font-weight:bold">BBa_K3809006: PuDHT</span></span></span></p></div><div id="comp-kuz0cnc9" class="XUUsC" title=""><div data-testid="linkElement" class="xQ_iF"><wix-image id="img_comp-kuz0cnc9" class="_1-6YJ _1Fe8-" data-image-info="{"containerId":"comp-kuz0cnc9","displayMode":"fill","imageData":{"width":936,"height":124,"uri":"d211b2_d63a5f8581b1425cb8167080d868b760~mv2.png","name":"T--TecCEM--fig5.png","displayMode":"fill"}}" data-bg-effect-name="" data-is-svg="false" data-is-svg-mask="false" data-image-zoomed="" data-has-ssr-src="" data-src="https://static.wixstatic.com/media/d211b2_d63a5f8581b1425cb8167080d868b760~mv2.png/v1/fill/w_750,h_99,al_c,q_85,usm_0.66_1.00_0.01/T--TecCEM--fig5.webp"><img src="https://static.igem.org/mediawiki/2021/7/74/T--TecCEM--fig5.png" alt="T--TecCEM--fig5.png" style="width:600px;height:79px;object-fit:cover;object-position:50% 50%"></wix-image></div></div><div id="comp-kuyzyyso2" class="_1Q9if" data-testid="richTextElement"><p class="font_8" style="font-size:18px; text-align:center"><span style="font-size:18px"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold"><span style="font-weight:bold">Figure 5.</span></span></span></p></div><div id="comp-kuyzyysq" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:17px; text-align:center"><span style="font-size:17px"><span class="color_15"><span style="font-family:wfont_069f35_23de2075afcc42cc848bac4f05c000f8,wf_23de2075afcc42cc848bac4f0,orig_roboto">Map of <span style="text-decoration:underline;"><a href="http://parts.igem.org/Part:BBa_K3809006" target="_blank" rel="noreferrer noopener">BBa_K3809006</a></span>. Created with <span style="text-decoration:underline"><a href="https://www.snapgene.com/" target="_blank" rel="noreferrer noopener"><span style="font-weight:bold">Snapgene</span></a></span></span></span></span></h2></div><div id="comp-kuyzyysr" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:20px; text-align:justify"><span class="color_15"><span style="font-size:20px"><span style="font-family:wfont_2ce676_53d594d91a1845238779d39b6e80ae45,wf_53d594d91a1845238779d39b6,orig_nunito_extralight">This sequence codes for the enzyme dehydratase from the organism Paralcaligenes ureilyticus. It was designed as part of a novel selection method in which the enzyme converts D-glyceric acid into Pyruvate, which is a useful compound for bacteria.</span></span></span></h2></div></div></div></div></div></section></div></div></div></div></section><section id="comp-kuz0cub2" class="_3d64y"><div id="bgLayers_comp-kuz0cub2" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuz0cub2" class="_2GUhU"></div></div><div data-testid="columns" class="_1uldx"><div id="comp-kuz0cudm" class="_1vNJf"><div id="bgLayers_comp-kuz0cudm" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuz0cudm" class="_2GUhU"></div></div><div data-mesh-id="comp-kuz0cudminlineContent" data-testid="inline-content" class=""><div data-mesh-id="comp-kuz0cudminlineContent-gridContainer" data-testid="mesh-container-content"><div id="comp-kuz0cudt" class="_1Q9if" data-testid="richTextElement"><h1 class="font_0" style="font-size:33px; text-align:center"><span style="color:#FAFAFA"><span style="font-size:33px"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold">BBa_K3809007: PuDHT + gldA homology regions</span></span></span></h1></div><div id="comp-kuz0fiwo" class="XUUsC" title=""><div data-testid="linkElement" class="xQ_iF"><wix-image id="img_comp-kuz0fiwo" class="_1-6YJ _1Fe8-" data-image-info="{"containerId":"comp-kuz0fiwo","displayMode":"fill","imageData":{"width":936,"height":130,"uri":"d211b2_2c920ae5ad6f424881435ac7c122ae82~mv2.png","name":"T--TecCEM--fig6.png","displayMode":"fill"}}" data-bg-effect-name="" data-is-svg="false" data-is-svg-mask="false" data-image-zoomed="" data-has-ssr-src="" data-src="https://static.wixstatic.com/media/d211b2_2c920ae5ad6f424881435ac7c122ae82~mv2.png/v1/fill/w_750,h_104,al_c,q_85,usm_0.66_1.00_0.01/T--TecCEM--fig6.webp"><img src="https://static.igem.org/mediawiki/2021/7/78/T--TecCEM--fig6.png" alt="T--TecCEM--fig6.png" style="width:600px;height:83px;object-fit:cover;object-position:50% 50%"></wix-image></div></div><div id="comp-kuz0cue3" class="_1Q9if" data-testid="richTextElement"><p class="font_8" style="font-size:18px; text-align:center"><span style="color:#FFFFFF"><span style="font-size:18px"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold"><span style="font-weight:bold">Figure 6.</span></span></span></span></p></div><div id="comp-kuz0cue5" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:17px; text-align:center"><span style="color:#FFFFFF"><span style="font-size:17px"><span style="font-family:wfont_069f35_23de2075afcc42cc848bac4f05c000f8,wf_23de2075afcc42cc848bac4f0,orig_roboto"> Map of <span style="text-decoration:underline;"><a href="http://parts.igem.org/Part:BBa_K3809007" target="_blank" rel="noreferrer noopener">BBa_K3809007</a></span>. Created with <span style="text-decoration:underline"><a href="https://www.snapgene.com/" target="_blank" rel="noreferrer noopener"><span style="font-weight:bold">Snapgene</span></a></span></span></span></span></h2></div><div id="comp-kuz0cue62" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:20px; text-align:justify"><span style="color:#FFFFFF"><span style="font-size:20px"><span style="font-family:wfont_2ce676_53d594d91a1845238779d39b6e80ae45,wf_53d594d91a1845238779d39b6,orig_nunito_extralight">This part corresponds to the coding sequence of alditol oxidase. It is regulated by the BBa_J23100 promoter and has a BBa_B0030 RBS and an rrnB T1 terminator BBa_B0010. It also has a spacer between the promoter and the RBS (BBa_B0040). The sequence contains two homology regions for the gene glycerol kinase (glpK) of <span style="font-style:italic;">E. coli,</span> which is a key enzyme in the glycerol metabolic pathway.</span></span></span></h2></div></div></div></div></div></section><section id="comp-kuz0cuea" class="_3d64y"><div id="bgLayers_comp-kuz0cuea" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuz0cuea" class="_2GUhU"></div></div><div data-testid="columns" class="_1uldx"><div id="comp-kuz0cued" class="_1vNJf"><div id="bgLayers_comp-kuz0cued" data-hook="bgLayers" class="_3wnIc"><div data-testid="colorUnderlay" class="_3KzuS _3SQN-"></div><div id="bgMedia_comp-kuz0cued" class="_2GUhU"></div></div><div data-mesh-id="comp-kuz0cuedinlineContent" data-testid="inline-content" class=""><div data-mesh-id="comp-kuz0cuedinlineContent-gridContainer" data-testid="mesh-container-content"><div id="comp-kuz0cuef" class="_1Q9if" data-testid="richTextElement"><p class="font_8" style="font-size:31px; text-align:center"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold"><span style="font-size:31px"><span style="font-weight:bold">BBa_K3809014: mRFP + oxyR homology regions + FLP recognition sequence</span></span></span></p></div><div id="comp-kuz0kgez" class="XUUsC" title=""><div data-testid="linkElement" class="xQ_iF"><wix-image id="img_comp-kuz0kgez" class="_1-6YJ _1Fe8-" data-image-info="{"containerId":"comp-kuz0kgez","displayMode":"fill","imageData":{"width":936,"height":128,"uri":"d211b2_03cf51da32be4af4af9af8769f598038~mv2.png","name":"T--TecCEM--fig7.png","displayMode":"fill"}}" data-bg-effect-name="" data-is-svg="false" data-is-svg-mask="false" data-image-zoomed="" data-has-ssr-src="" data-src="https://static.wixstatic.com/media/d211b2_03cf51da32be4af4af9af8769f598038~mv2.png/v1/fill/w_750,h_104,al_c,q_85,usm_0.66_1.00_0.01/T--TecCEM--fig7.webp"><img src="https://static.igem.org/mediawiki/2021/3/35/T--TecCEM--fig7.png" alt="T--TecCEM--fig7.png" style="width:600px;height:83px;object-fit:cover;object-position:50% 50%"></wix-image></div></div><div id="comp-kuz0cuei" class="_1Q9if" data-testid="richTextElement"><p class="font_8" style="font-size:18px; text-align:center"><span style="font-size:18px"><span style="font-family:wfont_299caa_3396eff86734448f84bd1bfa8335f847,wf_3396eff86734448f84bd1bfa8,orig_nunito_bold"><span style="font-weight:bold">Figure 7.</span></span></span></p></div><div id="comp-kuz0cuek" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:17px; text-align:center"><span style="font-size:17px"><span class="color_15"><span style="font-family:wfont_069f35_23de2075afcc42cc848bac4f05c000f8,wf_23de2075afcc42cc848bac4f0,orig_roboto">Map of <span style="text-decoration:underline;"><a href="http://parts.igem.org/Part:BBa_K3809014" target="_blank" rel="noreferrer noopener">BBa_K3809014</a></span>. Created with <span style="text-decoration:underline"><a href="https://www.snapgene.com/" target="_blank" rel="noreferrer noopener"><span style="font-weight:bold">Snapgene</span></a></span></span></span></span></h2></div><div id="comp-kuz0cuen" class="_1Q9if" data-testid="richTextElement"><h2 class="font_2" style="font-size:20px; text-align:justify"><span class="color_15"><span style="font-size:20px"><span style="font-family:wfont_2ce676_53d594d91a1845238779d39b6e80ae45,wf_53d594d91a1845238779d39b6,orig_nunito_extralight">This sequence codes for the reporter protein mRFP. It was designed as part of a novel selection method in which the oxidative stress regulator gene (oxyR) is removed. mRFP will act as a reporter and will indicate if the knockout was successful or not. This gene will work alongside aldO, PuDHT and katE to develop a new selection mechanism which involves the knockout of genes gldA, glpK and oxyR to make <span style="font-style:italic;">E. coli </span>susceptible to death by exposure to low concentrations of glycerol.</span></span></span></h2></div></div></div></div></div></section><div data-mesh-id="Containertv0y0inlineContent-wedge-9"></div></div></div></div></div></div></div></div></main> | |
Revision as of 05:09, 20 October 2021
Home
Team
Project
Awards
Human Practices
Parts
Judging form
Selection Marker Parts
Biobricks to revolutionize the Safety in the Lab
The design of these parts was made thinking about a novel selection marker and a modified strain of E. coli we will use to substitute the use of antibiotics in genetic engineering protocols. The idea is to use this parts to generate a knockout strain from E. coli DH5⍺ genes oxyR, gldA and glpK and substitute with aldO, mRFP and PuDHT genes. We also designed a part that forms a plasmid and acts as a selection cassette. AldO will convert glycerol to peroxide and D-glyceric acid, katE will convert peroxide to water and oxygen and PuDHT will convert D-glyceric acid into pyruvate. We will briefly explore the functionality of each part, as well as key aspects of its design.
BBa_K3809001: Novel Selection Cassette katE
Figure 1.
Map of BBa_K3809001. Created with Snapgene.
This part codes for the E. coli catalase gene katE which converts peroxide into water and oxygen. It is meant to be used with a pUC backbone we designed in which the part acts as a selection marker for the modified strain. For more information, click on the part name.
BBa_K3809003: katE regulated by BBa_J23100
Figure 2.
Map of BBa_K3809003. Created with Snapgene
This Biobrick contains a katE gene with an RBS (BBa_B0030), spacer (BBa_B0040), terminator (BBa_B0010) and promoter (BBa_J23100). It was designed to be expressed as part of a plasmid, to act as a selection cassette for a modified strain of E. coli.
BBa_K3809004: aldO
Figure 3.
Map of BBa_K3809004. Created with Snapgene.
This sequence codes for the enzyme alditol oxidase from the organism Streptomyces coelicolor. It was designed as part of a novel selection method in which the enzyme converts glycerol into D-glycerate.
BBa_K3809005: aldO + glpK homology regions
Figure 4.
Map of BBa_K3809005. Created with Snapgene
This part corresponds to the coding sequence of alditol oxidase. It is regulated by the BBa_J23100 promoter and has a BBa_B0030 RBS and an rrnB T1 terminator BBa_B0010. It also has a spacer between the promoter and the RBS (BBa_B0040). The sequence contains two homology regions for the gene glycerol kinase (glpK) of E. coli, which is a key enzyme in the glycerol metabolic pathway.
BBa_K3809006: PuDHT
Figure 5.
Map of BBa_K3809006. Created with Snapgene
This sequence codes for the enzyme dehydratase from the organism Paralcaligenes ureilyticus. It was designed as part of a novel selection method in which the enzyme converts D-glyceric acid into Pyruvate, which is a useful compound for bacteria.
BBa_K3809007: PuDHT + gldA homology regions
Figure 6.
Map of BBa_K3809007. Created with Snapgene
This part corresponds to the coding sequence of alditol oxidase. It is regulated by the BBa_J23100 promoter and has a BBa_B0030 RBS and an rrnB T1 terminator BBa_B0010. It also has a spacer between the promoter and the RBS (BBa_B0040). The sequence contains two homology regions for the gene glycerol kinase (glpK) of E. coli, which is a key enzyme in the glycerol metabolic pathway.
BBa_K3809014: mRFP + oxyR homology regions + FLP recognition sequence
Figure 7.
Map of BBa_K3809014. Created with Snapgene
This sequence codes for the reporter protein mRFP. It was designed as part of a novel selection method in which the oxidative stress regulator gene (oxyR) is removed. mRFP will act as a reporter and will indicate if the knockout was successful or not. This gene will work alongside aldO, PuDHT and katE to develop a new selection mechanism which involves the knockout of genes gldA, glpK and oxyR to make E. coli susceptible to death by exposure to low concentrations of glycerol.
-
Index
Tips
Hover the references