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<img src="https://static.igem.org/mediawiki/2016/0/05/T--Edinburgh_OG--Matin_Back_Interlab.jpg" width= "95%"> | <img src="https://static.igem.org/mediawiki/2016/0/05/T--Edinburgh_OG--Matin_Back_Interlab.jpg" width= "95%"> | ||
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<br><mark>Thus, we explore suitable organisms for the tasks and develop the means to do genetic manipulation and create a collection of necessary parts library.</mark> <br>In the process, we realized that by domesticating new organisms as chassis, we open up new risks and safety challenges towards users and environment. | <br><mark>Thus, we explore suitable organisms for the tasks and develop the means to do genetic manipulation and create a collection of necessary parts library.</mark> <br>In the process, we realized that by domesticating new organisms as chassis, we open up new risks and safety challenges towards users and environment. | ||
<br><mark>Therefore, we also developed a tool (software) to screen for potential risk from the organism’s database of toxic secondary metabolites.</mark> | <br><mark>Therefore, we also developed a tool (software) to screen for potential risk from the organism’s database of toxic secondary metabolites.</mark> | ||
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+ | <center><a href="https://static.igem.org/mediawiki/2016/0/08/T--Edinburgh_OG--Tom_Essay_WhyNeedMoreChassis.pdf" class="page-scroll btn btn-default btn-xl sr-button">An Essay on Expanding Chassis</a> | ||
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<p><b>Rhodococcus jostii</b> is a gram positive bacteria with extensive catabolic pathway to degrade variety of chlorinated compounds, such as polychlorinated biphenyls (PCBs). They are potential chassis for synthetic biology applications in bioremediation.</p> | <p><b>Rhodococcus jostii</b> is a gram positive bacteria with extensive catabolic pathway to degrade variety of chlorinated compounds, such as polychlorinated biphenyls (PCBs). They are potential chassis for synthetic biology applications in bioremediation.</p> | ||
<p><b>Penicillium roqueforti</b> is a filamentous fungi used in the production of blue cheese. They are biotechnologically relevant for the industrial production of enzymes (cellulases, pectinases, lipases, proteases and amylases) and could be used to produce complex secondary metabolites.</p> | <p><b>Penicillium roqueforti</b> is a filamentous fungi used in the production of blue cheese. They are biotechnologically relevant for the industrial production of enzymes (cellulases, pectinases, lipases, proteases and amylases) and could be used to produce complex secondary metabolites.</p> | ||
− | </div> | + | <center><a href="https://2016.igem.org/Team:Edinburgh_OG/Experiments" class="page-scroll btn btn-default btn-xl sr-button">Find out more!</a> |
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<br><br>To utilise MoClo, a set of destination vectors need to be developed to accommodate assembly in different levels. Unfortunately, the chassis we were working on (R. jostii) cannot utilise the origin of replication from available destination vectors, which are designed for Eschericia coli. Therefore, we developed a set of MoClo destination vectors for assembly and transformation to R. jostii. We have shown that our destination vectors can be used for combinatorial assembly and transformed to the host organism. | <br><br>To utilise MoClo, a set of destination vectors need to be developed to accommodate assembly in different levels. Unfortunately, the chassis we were working on (R. jostii) cannot utilise the origin of replication from available destination vectors, which are designed for Eschericia coli. Therefore, we developed a set of MoClo destination vectors for assembly and transformation to R. jostii. We have shown that our destination vectors can be used for combinatorial assembly and transformed to the host organism. | ||
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+ | <a href="https://2016.igem.org/Team:Edinburgh_OG/Description#design" class="page-scroll btn btn-default btn-xl sr-button">Vector Design</a> | ||
+ | <a href="https://2016.igem.org/Team:Edinburgh_OG/Proof" class="page-scroll btn btn-default btn-xl sr-button">Proof of Concept</a> | ||
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<p class="text-faded" style="font-size: 15px">To utilise synthetic biology, a working parts (promoters, RBS, coding sequences) need to be developed as not all of them works orthogonally. We have developed a set of useful parts for expression in our chassis. | <p class="text-faded" style="font-size: 15px">To utilise synthetic biology, a working parts (promoters, RBS, coding sequences) need to be developed as not all of them works orthogonally. We have developed a set of useful parts for expression in our chassis. | ||
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− | </div> | + | <center><a href="https://2016.igem.org/Team:Edinburgh_OG/Parts" class="page-scroll btn btn-default btn-xl sr-button">Find out more!</a> |
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<h3>Human Practices<h3> | <h3>Human Practices<h3> | ||
<p class="text-faded" style="font-size: 15px">The domestication of non-model organisms raises both biosafety and biosecurity challenges (e.g. unknown pathogenicity and toxicity). Such issues, if are not properly addressed, can be a risk for both users and their surroundings. The Edinburgh OG team worked to develop an accessible, easy-to-use program to evaluate the toxicity of curated secondary metabolites from organisms used as chassis. We have incorporate this aspect in our project and communicate with other teams in order to give contribution towards current risk assessment procedures and as precautionary step for both experienced and non-experienced users. | <p class="text-faded" style="font-size: 15px">The domestication of non-model organisms raises both biosafety and biosecurity challenges (e.g. unknown pathogenicity and toxicity). Such issues, if are not properly addressed, can be a risk for both users and their surroundings. The Edinburgh OG team worked to develop an accessible, easy-to-use program to evaluate the toxicity of curated secondary metabolites from organisms used as chassis. We have incorporate this aspect in our project and communicate with other teams in order to give contribution towards current risk assessment procedures and as precautionary step for both experienced and non-experienced users. | ||
</p> | </p> | ||
+ | <center><a href="https://2016.igem.org/Team:Edinburgh_OG/Description#design" class="page-scroll btn btn-default btn-xl sr-button">Find out more!</a> | ||
+ | </center></div> | ||
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<figcaption>Fig1. (A) Physical map of pSRKBB-Empty showing pSR1 replicon as a black band. The replicon contains per (a positive effector of replication) and rep (replicase) gene. pK19 backbone (white band) contains pBR322 ORI and Kanamycin resistance gene (KmR) regulated by the P45 promoter. Mutagenic primer binding sites are shown as red arrows (mut1, mut2, mut3, and mut4). The region in the red box was replaced by (B) a BioBrick cloning site flanked by standard verification primer binding sites, VF2 and VR, derived from Iverson et al. (2016), creating pSRKM_Empty. Destination vectors can be made by digestion and ligation with SpeI to insert standardized LacZα flanked by the designated Type IIs restriction enzyme (recognition sites shown as bold) and four base overhangs for Level-1 (C) or Level-2 (D) assembly. (E) Schematic map of the level-1 destination vector pSRKM_NN and (F) the level-2 destination vector pSRCM_NN with chloramphenicol resistance (CmR). X refers to the fusion sites and NNNN to its four base overhangs: A (GGAG), B (TACT), C (AATG), D (AGGT), E (GCTT), F (CGCT), G (TGCC), and H (ACTA).</figcaption> | <figcaption>Fig1. (A) Physical map of pSRKBB-Empty showing pSR1 replicon as a black band. The replicon contains per (a positive effector of replication) and rep (replicase) gene. pK19 backbone (white band) contains pBR322 ORI and Kanamycin resistance gene (KmR) regulated by the P45 promoter. Mutagenic primer binding sites are shown as red arrows (mut1, mut2, mut3, and mut4). The region in the red box was replaced by (B) a BioBrick cloning site flanked by standard verification primer binding sites, VF2 and VR, derived from Iverson et al. (2016), creating pSRKM_Empty. Destination vectors can be made by digestion and ligation with SpeI to insert standardized LacZα flanked by the designated Type IIs restriction enzyme (recognition sites shown as bold) and four base overhangs for Level-1 (C) or Level-2 (D) assembly. (E) Schematic map of the level-1 destination vector pSRKM_NN and (F) the level-2 destination vector pSRCM_NN with chloramphenicol resistance (CmR). X refers to the fusion sites and NNNN to its four base overhangs: A (GGAG), B (TACT), C (AATG), D (AGGT), E (GCTT), F (CGCT), G (TGCC), and H (ACTA).</figcaption> | ||
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Latest revision as of 19:52, 19 October 2016
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