Team:NCTU Formosa/Project
From 2012.igem.org
Introduction to the project
(developing)
Project details
Enzyme for isobutanol
(developing)
Temperature control system
The low temperature release system is a way to let e.coli produce isobutanol efficiently . Because isobutanol and isobutyaldehyde are toxic to the e.coli , the system avoid e.coli facing them at the beginning . The following picture is our system.
At the beginning , we will let E.coli stay in 37°C environment. After having enough 2-Ketoisovalerate , we will move E.coli into 30°C environment for producing the final product , isobutanol. It can make us get isobutanol successfully and efficiently.
This is our biobrick. The most important part of our biobrick is 37°C ribosome binding site gene. We separate our biobrick into two parts . The first part is which has 37℃ ribosome binding site gene and the second part is under the first one.
And now we're introducing how our system works.
When being in 37°C environment, the first part will be translated and produce tetR protein to inhibit Ptet promoter. The second part will not be translated. Then we can produce intermediate , 2-Ketoisovalerate.
After getting enough 2-Ketoisovalerate , E.coli will stay in 30°C environment. The ribosome will not bind the 37°C ribosome binding site and tetR genes will not be translated. Then the second part will be translated successfully. At the end , we can get the isobutanol.
Result
The report shows that we use fluorescene protein to mark the second part of our biobrick . We can see that fluorescene protein staying in 30°C environment will have higher expression than staying in 37°C environment . According to it , we can see that our system do truly work!
Zinc finger
(developing)
Result
(developing)
Instrument
(developing)
Conclusion
(developing)