Experiment
Our product aims to reach a high quality of “long-term monitor”, which means considerably stable expression of color is needed. Thus, we choose chromoprotein as our reporter protein due to its steadily expression and low half-life degradation rate. (See more in here)
cj-Blue
We add a LVA tag to cjBlue in order to accelerate its degrade speed. (See more in here)
Concentration simulation of cjBlue
Chromoprotein
According to our model of chromoprotein
Expression
Temperature
35ﾟC / 37ﾟC / 39ﾟC / 32ﾟC
Chromoprotein o.d. value
(1) RFP (E1010): 607nm
(2) BFP (k592009):588nm
(3) cjBlue (k592011):610nm
Culture Setting
(1) 32ﾟC, 200rpm, culture12hr (15ml) *continue to log phase
(2) add (15ml) LB *re-culture
(3) TeﾟC, continue cultural
(4) Take samples from the centrifuge tube per hour, and narrow the time range and improve the accuracy.
UV
Culture setting
(1) 37ﾟC, 200rpm, culture 12hr (15ml) *continue to log phase
(2) Add (15ml) LB *re-culture
(3) re-culture at 37ﾟC under UV
(4) Take samples from the centrifuge tube per hour, and narrow the time range and improve the accuracy.
Expression measurement setting
(1) Extract: Take (3ml) bacteria liquid, and centrifuge at 14000rpm, 1min
(2) add 1ml PBS, pipetting+vortex
(3) measure the o.d. value
