Firstly we made a basic part of magainin-2 sequense, toxic protein that derives from Xenopus laevis. Secondly, we made its expression visible by making a composite part that expresses GFP with magainin-2.
Initially we were strongly inspired by the project of UT-Tokyo2016,color-changing E.coli dependently on cell divisions and determined to develop their project. Our current project is kind of a section of the initial project which was too content-full for a year.
Our Initial Project
Generation 1 RED
Generation 2 BLUE
Generation 3 GREEN
Generation 4 DEATH!!
Designed by team UT-Tokyo 2016, cited from
1. BBa_K3050000 (Partially validated)
This is a composite part that includes T7 promotor + RBS + mag-2 + GFP + double terminators.
BBa_I746909 is linerized by Inverse PCR to be combined with magainin-2 sequense by In-Fusion reaction.
BL21(DE3) transformed pSB1C3 BBa_K3050000 successfully grew and emit strong green fluorescent light as shown in pictures.
Thus we succeeded to prove Inverse PCR,In-Fusion reaction,and transformation went great.
GFP part is on downstream of T7 promotor and magainin-2, hence we're quite sure that this result indicates that magainin-2 is expressed at the same time.
Fig.1 BL21(DE3) pSB1C3 BBa_K3050000 expressed under visible light with negative control without IPTG induction
BL21(DE3) pSB1C3 BBa_K3050000, under visible light, 3.5 hours after directly added 300 μM IPTG.
Fig.2 pSB1C3 BBa_K3050000 expressed under UV light with negative control without IPTG induction
BL21(DE3) pSB1C3 BBa_K3050000, under UV light, 3.5 hours after directly added 300 μM IPTG.
Fig.3 Four biological replicates all highly expressed
BL21(DE3) pSB1C3 BBa_K3050000, under visible light, 26 hours after directly added 500 μM IPTG.
The fluorescent light didn't extingished for more than whole a day after added 500 μM IPTG.
2. BBa_K3050001 (Validated as a part of BBa_K3050000)