Difference between revisions of "Part:BBa K3788020:Experience"
(→Applications of BBa_K3788020) |
|||
| Line 4: | Line 4: | ||
how you used this part and how it worked out. | how you used this part and how it worked out. | ||
| − | === | + | |
| + | |||
| + | <h1>Applications of BBa_K3788020</h1> | ||
| + | ===Method=== | ||
| + | <p>In order to study the previously reported latency period between induction of the <i>caa-cal</i> operon system, ColicinA production, and CaL lysis protein production, we fused the RFP coding sequence (BBa_E1010) with the Modified lysis device part (BBa_K3788015) (composite part BBa_K3788020). Induction of the SOS response was induced by mitomycin C treatments and fluorescence was recorded as a function of dose using a microplate reader.</p> | ||
| + | |||
| + | ===Results=== | ||
| + | <p>Cells carrying the BBa_K3788020 construct were cultured in the presence of various concentrations of mitomycin C, ranging from 0 to 800 ng.ml-1. The OD (600 nm) and the amount of red fluorescence were recorded over time.</p> | ||
| + | |||
| + | https://2021.igem.org/wiki/images/2/26/T--Aix-Marseille--The_timer_effect_delayed_synthesis_of_the_cal_RFP_protein_production.jpeg | ||
| + | |||
| + | <p>We observed a strong dose-dependent synthesis of our reporter protein during treatment with mitomycin C. The signal was detected after 330 min to 420 min of induction, for the highest dose and the lowest dose, respectively.</p> | ||
| + | |||
| + | <p>This contrasts with what we observed when GFP was under the control of the LexA-dependent promoter (BBa_K3780817) where we could observe the accumulation of green fluorescence as early as 40 minutes after induction.</p> | ||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 23:57, 21 October 2021
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K3788020
Method
In order to study the previously reported latency period between induction of the caa-cal operon system, ColicinA production, and CaL lysis protein production, we fused the RFP coding sequence (BBa_E1010) with the Modified lysis device part (BBa_K3788015) (composite part BBa_K3788020). Induction of the SOS response was induced by mitomycin C treatments and fluorescence was recorded as a function of dose using a microplate reader.
Results
Cells carrying the BBa_K3788020 construct were cultured in the presence of various concentrations of mitomycin C, ranging from 0 to 800 ng.ml-1. The OD (600 nm) and the amount of red fluorescence were recorded over time.
We observed a strong dose-dependent synthesis of our reporter protein during treatment with mitomycin C. The signal was detected after 330 min to 420 min of induction, for the highest dose and the lowest dose, respectively.
This contrasts with what we observed when GFP was under the control of the LexA-dependent promoter (BBa_K3780817) where we could observe the accumulation of green fluorescence as early as 40 minutes after induction.
User Reviews
UNIQ1e92554085477bc3-partinfo-00000000-QINU UNIQ1e92554085477bc3-partinfo-00000001-QINU