Team:SYSU-China/Project

From 2014.igem.org

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<p>Tell us more about your project.  Give us background.  Use this as the abstract of your project.  Be descriptive but concise (1-2 paragraphs)</p>
<p>Tell us more about your project.  Give us background.  Use this as the abstract of your project.  Be descriptive but concise (1-2 paragraphs)</p>
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<p>The contemporary methods of directed evolution of certain protein require much work by people. This summer, SYSU-China is trying to construct an in vitro system which can allow us to directly obtain a particular protein in a simple and convenient way. The key of this system is to turn protein interaction signal into the advantage of evolution. We chose the bacteria two hybrid system to transfer the protein interaction signal into the expression of a target gene, gene II from M13 bacteria phage, which is a key factor for phage DNA replication. By knocking out the gene II in M13 bacteria phage genome and expressing gene II under the control of the bacteria two hybrid system, we can successfully convert the protein interaction signal into the advantage of M13 phage generation. The M13 phage generated in our system carry the protein coding sequence needed to evolve. As the system operating, the phage contained desirable protein coding sequence can grow into dominant population.</p>
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<p>Current methods of high-affinity protein selection are time-consuming and laborious, which hinders the application process. This year, by constructing an integrated protein evolution machine, we made it automatic and efficient. A broad diversification was accomplished by a DNA mutagenesis module in the host bacteria, which enabled prototype protein sequence, carried by the budding-deficient M13 bacteriophage, to generate a library of candidate proteins. Subsequently, bacterial-two-hybrid system was used to give selection pressure on these candidates. Only the favorable candidate proteins can activate compensating gene transcription and rescue phage budding, thus enriching favorable protein sequence in the phage population. Moreover, an RNA thermometer, on the compensating gene mRNA, was applied in the phage amplification process to make it more controllable. Finally, by integrating all the diversification, selection and amplification process, our integrated directed evolution machine can make it an automatic and controllable process for high-affinity protein selection. </p>
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<p>What’s more, in order to make this system more efficient and more controllable, we introduce several proteins to raise the mutation rate and use RNA thermometer to control gene II expression in the dimension of temperature.</p>
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<p>This system can have broad applications. It may be able the generate antibody that we cannot obtain in normal way, for example, the HIV antibody. We can integrate HIV antigen into our system and generate the corresponding antibody. </p>
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<h3>References </h3>
<h3>References </h3>

Latest revision as of 22:38, 16 October 2014



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Project Description

Content

Tell us more about your project. Give us background. Use this as the abstract of your project. Be descriptive but concise (1-2 paragraphs)

Current methods of high-affinity protein selection are time-consuming and laborious, which hinders the application process. This year, by constructing an integrated protein evolution machine, we made it automatic and efficient. A broad diversification was accomplished by a DNA mutagenesis module in the host bacteria, which enabled prototype protein sequence, carried by the budding-deficient M13 bacteriophage, to generate a library of candidate proteins. Subsequently, bacterial-two-hybrid system was used to give selection pressure on these candidates. Only the favorable candidate proteins can activate compensating gene transcription and rescue phage budding, thus enriching favorable protein sequence in the phage population. Moreover, an RNA thermometer, on the compensating gene mRNA, was applied in the phage amplification process to make it more controllable. Finally, by integrating all the diversification, selection and amplification process, our integrated directed evolution machine can make it an automatic and controllable process for high-affinity protein selection.


References

iGEM teams are encouraged to record references you use during the course of your research. They should be posted somewhere on your wiki so that judges and other visitors can see how you though about your project and what works inspired you.

You can use these subtopics to further explain your project

  1. Overall project summary
  2. Project Details
  3. Materials and Methods
  4. The Experiments
  5. Results
  6. Data analysis
  7. Conclusions

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