Project

Circular mRNA -the world´s longest protein-

1. Overall project summary
2. Introduction
3. Project Flow
4. Theory and Methods
5. Experiments
6. Results&Data analysis
7. Future Works
8. Conclusions
9. References

Project Description

This year, the theme of iGEM Gifu is to synthesize a long chain protein with circular mRNA. It is a sequel to our theme of the last year.

Here, we explain our theme of the last year.

The circular mRNA is expressed in E.coli by using a mechanism of self-splicing.

In td gene of T4 phage, there is the group 1 intron. A part of the RNA transcribed from the group 1 intron is a ribozyme which catalyzes a splicing reaction, and this RNA splices itself.

The circular mRNA can be expressed in E.coli by cloning a domain which functions as ribozyme for splicing of the td gene, and putting it in the plasmid. This method is called “PIE method.”

The circular mRNA without the termination codon sequence can be used to synthesize protein semi-permanently.

We succeeded in expression of the circular mRNA, and synthesizing long chain protein last year. But the long chain protein synthesized in last year was not functional because its holding had lost shape. So, this year, we design a linker sequence and synthesize a functional long chain protein. On the other hand, the probability was just a few percent that the cyclization happens. It is thought that the ribozyme site of the splicing is far from the reactive site, and it lowers the probability. So we’ll design complemental sequence near the ribozyme site and the reactive site of splicing to make them accessible to each other, and raise the probability of the cyclization in the theme.

Before you start:

Please read the following pages:

• Requirements page
• Wiki Requirements page

Editing your wiki

On this page you can document your project, introduce your team members, document your progress and share your iGEM experience with the rest of the world!

Click here to edit this page!

See tips on how to edit your wiki on the Template Documentation page.

Templates

This year we have created templates for teams to use freely. More information on how to use and edit the templates can be found on the Template Documentation page.

Tips

This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started:

• State your accomplishments! Tell people what you have achieved from the start.
• Be clear about what you are doing and how you plan to do this.
• You have a global audience! Consider the different backgrounds that your users come from.
• Make sure information is easy to find; nothing should be more than 3 clicks away.
• Avoid using very small fonts and low contrast colors; information should be easy to read.
• Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the iGEM 2015 calendar
• Have lots of fun!

Inspiration

You can also view other team wikis for inspiration! Here are some examples:

• 2014 SDU Denmark
• 2014 Aalto-Helsinki
• 2014 LMU-Munich
• 2014 Michigan
• 2014 ITESM-Guadalajara
• 2014 SCU-China

Uploading pictures and files

You can upload your pictures and files to the iGEM 2015 server. Remember to keep all your pictures and files within your team's namespace or at least include your team's name in the file name.
When you upload, set the "Destination Filename" to Team:YourOfficialTeamName/NameOfFile.jpg. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!)