Difference between revisions of "Team:Peking/Demonstrate"

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                             <p><b>We summarized the advantages of the Uranium Reaper system in Table 1.</b></p>
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                             <p>Even though the efficiency of the Uranium Reaper system may be somewhat lower than current methods, it can certainly be optimized through further development work. Importantly, Uranium Reaper is much better in other aspects. In the future, we plan to optimize the entire Uranium Reaper strategy in order to enhance the adsorption efficiency.</p>
 
                             <p>Even though the efficiency of the Uranium Reaper system may be somewhat lower than current methods, it can certainly be optimized through further development work. Importantly, Uranium Reaper is much better in other aspects. In the future, we plan to optimize the entire Uranium Reaper strategy in order to enhance the adsorption efficiency.</p>
 
                              
 
                              
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             <div class="texttitle" style="margin-top:40px;">Our future plan</div>
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             <div class="texttitle" style="margin-top:40px;">Our future plan</div>
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Revision as of 12:10, 15 October 2016

Recovery

Demonstration.

Project Achievement

1.

Constructed a multifunctional protein sequences of interest via molecular biological methods, and introduced the constructed plasmids into the engineered bacteria for protein expression. (Learn more)

2.

Searched for methods and the best conditions for the extraction of each protein. (Learn more)

3.

Demonstrated a quick and stable crosslinking process of Triple SpyTag-SUP and Triple SpyTag-mSA with Triple SpyCatcher via covalent bonds. We also optimized this reaction concerning the relevant parameters such as temperature, pH, etc., and confirmed that changes of pH or temperature would not interfere with the whole process. (Learn more)

4.

Demonstrated effective adsorption of uranyl ions by monomeric Triple SpyTag-SUP or protein networks containing the SUP module under a number of conditions. The adsorption was highly efficient and fast, not only under experimental conditions but also in simulated seawater or freshwater containing uranium pollution. (Learn more)

5.

Attached biotin to amino-coated magnetic beads and achieved retrieval of the hydrogel formed via the crosslinking of Triple SpyTag-SUP and Triple SpyTag-mSA with Triple SpyCatcher with a magnet. (Learn more)

6.

Set up a signal peptide library and screened for optimally suited signal peptides in order to efficiently secrete the proteins of interest. We found two signal peptides of high efficiency - those derived from OmpA and LtIIB. (Learn more)

7.

Used all the above-mentioned experiments together to demonstrate that the complete Uranium Reaper system, consisting of Triple SpyTag-SUP, Triple SpyTag-mSA, Triple SpyCatcher and biotin-coated magnetic beads, could effectively handle uranium pollution under simulated real-life conditions in about 2 hours. We aim to optimize this strategy and hope it can be implemented as a uranyl removal kit. (Learn more)

8.

We exchanged the Triple SpyTag-SUP monomer for Triple SpyTag-LBP or Triple SpyTag-CBP, and tried using the same strategy to adsorb lead and cadmium. The results were remarkable, clearly demonstrating that the Uranium Reaper strategy has much potential to be expanded to other heavy metals. (Learn more)


We summarized the advantages of the Uranium Reaper system in Table 1.

Even though the efficiency of the Uranium Reaper system may be somewhat lower than current methods, it can certainly be optimized through further development work. Importantly, Uranium Reaper is much better in other aspects. In the future, we plan to optimize the entire Uranium Reaper strategy in order to enhance the adsorption efficiency.

Beyond Experiment

1.

We submitted about 70 high-quality and well-characterized Standard BioBricks, including a set of derivatives of triple SpyTag and triple SpyCatcher, such as the triple SpyTag-SUP and triple SpyTag-mSA. (Learn more)

2.

We developed a special software which can be used to calculate the molecular weight distribution of protein polymers using Flory’s theory. The results of testing have demonstrated that the software is accurate and useful. (Learn more)

3.

We visited experts from the Peking University departments for Nuclear & Radiochemistry and Physics, respectively, to learn about the current situation surrounding uranium pollution in the real world and how people can control the situation. After finishing the main work, we presented them with the achievements of the project and got their feedback. (Learn more)

4.

We did an interview with the Hunan Nuclear Geology 311 Brigade and gained thorough insights into the treatment of uranyl pollution used by the people on the firing line. This way we could compare the methods they were using with the Uranium Reaper strategy. (Learn more)

5.

We helped and collaborated with 7 other iGEM teams by guiding a new team (BHU-China), as well as discussing about project design and technical skills and sharing DNA materials (OUC-China, BIT-China, Tianjin, UCAS, Jinlin-China and BNU-China). (Learn more)

6.

We attended the CCiC (Central China iGEM Consortium), which is a large-scale competition-free jamboree of about 50 teams, providing participants with an opportunity for meaningful exchanges of ideas and problem solving. (Learn more)

Our future plan

1.

We should reproduce all of the experiments that we have done this summer to make sure the results are credible.

2.

We will optimize the whole strategy to enhance the adsorption efficiency by changing pH, temperature, reaction time of crosslinking and recovery. (The efficiency is only about 60% without further optimization)

3.

According to the results for the adsorption of 13nM uranyl, the hydrogel exhibited a good ability in a simulated seawater environment. We can thus also look into other usage scenarios of Uranium Reaper, such as bio-mining and uranium enrichment.

4.

Exchange of the SUP module for other functional proteins. For example, we can integrate proteins which can bind other heavy metals such as mercury so that the hydrogel can be used to treat other kinds of pollution as well.

5.

We can assemble enzyme systems behind the SpyTag backbone to create a production plant in vitro. In the protein hydrogel, the concentration of enzymes can be increased and the efficiency of biocatalysis may consequently also be enhanced.

6.

If we optimize the number of SpyTag or SpyCatcher modules per protein monomer, as well as the working concentrations of proteins, we may make protein-3D printing using the Spy Crosslinking Network come true.