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ncAA Kit

Content

Noncanonical amino acids (ncAAs) are an exciting new tool in the biological researchers toolbox. Incorporating ncAAs into proteins will allow scientists to create bacteria that can perform novel or new functions. The amber stop codon has been recoded and removed from the E. coli genome in order to be used as the codon for ncAAs. Unfortunately, these unnatural amino acids are often difficult to incorporate into proteins due to a low fidelity of the synthetase/tRNA pair. Our project aims to create a kit that can measure the fidelity of the synthetase/tRNA pair and incorporation of several different unnatural amino acids into fluorescent proteins. The construct for the kit is a simple 2 plasmid system. The first plasmid is IPTG inducible and contains RFP, followed by a linker sequence containing a recoded amber stop codon (where the ncAA will be incorporated) and sfGFP. The other plasmid contains the synthetase/tRNA pair. The cells can be induced with IPTG and the protein fusion can be illuminated. Depending on the relative intensities of the RFP and GFP fluorescent proteins, we can determine how efficient the synthetase/tRNA pairs are at incorporating the ncAA. We plan to equip researchers with a quick and easy “plug and play” system that contains easily interchangeable parts. Researchers will be able to insert any plasmid containing a new synthetase/tRNA pair into our pre-made cells to test the fidelity and incorporation of various ncAAs.

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

It's important for teams to describe all the creativity that goes into an iGEM project, along with all the great ideas your team will come up with over the course of your work.

It's also important to clearly describe your achievements so that judges will know what you tried to do and where you succeeded. Please write your project page such that what you achieved is easy to distinguish from what you attempted.

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)

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

It's important for teams to describe all the creativity that goes into an iGEM project, along with all the great ideas your team will come up with over the course of your work.

It's also important to clearly describe your achievements so that judges will know what you tried to do and where you succeeded. Please write your project page such that what you achieved is easy to distinguish from what you attempted.

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)

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

It's important for teams to describe all the creativity that goes into an iGEM project, along with all the great ideas your team will come up with over the course of your work.

It's also important to clearly describe your achievements so that judges will know what you tried to do and where you succeeded. Please write your project page such that what you achieved is easy to distinguish from what you attempted.