Introduction
In our experiment of the E.co-mail, the important thing is to degrade GFP-Lon-tag more rapidly. GFP is usually stable and has a long half–life. So, we thought of how to measure the reaction rate.
Method
E.coli (pIJ106b, pJT122, plPCB) is cultivated in TY culture medium for 7 hours. It was precipitated by centrifugation and washed with 1 % NaClaq.
400 μL of 1% NaCl was added to the E.coli sample and protein synthesis was stopped by addition of tetracyclin at the concentration of 100 μg/mL.
An aliquot from each sample was plated on agar and cultured overnight.
The solution was exposed to blue LED light, and we took a color picture every 1 hour for 6 hours.
By using the ImageJ software, we measured the intensity of light from GFP.
"
