Difference between revisions of "Team:KAIT JAPAN/Demonstrate"
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| + | <h1><font color="#3335FF">DEMONSTRATE</font></h1> | ||
| + | <p>  Functional test of this system was carried out by adding IL-10 to E. coli having plasmid of IL-12. And we will investigate whether IL-12 is transported out of the cell by performing SDS-page after collecting the media. And we will check the expression of IL-12 by western blotting.</p> | ||
| + | <p><font color="#FF3333">Detects and check whether IL-12 is transport out of the cell.</font></p> | ||
| + | <p><center><img src="https://static.igem.org/mediawiki/2017/9/9e/T--KAIT_JAPAN--function_IL-12.png"height="650" width="350"></center> | ||
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| + | <p>However, there is a big problem to conduct functional test by using this method. That is, IL-12 antibodies used for Western blotting is expensive. Therefore, we will use GFP to check whether the system works properly. IL-10 is added to the medium of E. coli carrying two plasmids (GFP) and observed whether GFP is expressed. When the expression of GFP on the medium is confirmed, we will then observe the medium with fluorescence microscope to confirm whether GFP has been transported out of the cell.</p> | ||
| + | <p><center><img src="https://static.igem.org/mediawiki/2017/1/14/T--KAIT_JAPAN--function_GFP.png"height="650" width="500"> | ||
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| + | <p>If the functional test using GFP succeeds, we will test this system at the protein level.</p> | ||
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Latest revision as of 20:39, 1 November 2017
DEMONSTRATE
Functional test of this system was carried out by adding IL-10 to E. coli having plasmid of IL-12. And we will investigate whether IL-12 is transported out of the cell by performing SDS-page after collecting the media. And we will check the expression of IL-12 by western blotting.
Detects and check whether IL-12 is transport out of the cell.
However, there is a big problem to conduct functional test by using this method. That is, IL-12 antibodies used for Western blotting is expensive. Therefore, we will use GFP to check whether the system works properly. IL-10 is added to the medium of E. coli carrying two plasmids (GFP) and observed whether GFP is expressed. When the expression of GFP on the medium is confirmed, we will then observe the medium with fluorescence microscope to confirm whether GFP has been transported out of the cell.
If the functional test using GFP succeeds, we will test this system at the protein level.
Contact: kaitjapan@gmail.com Twitter:@KAIT_JAPAN
