Team:NUDT CHINA/Composite Parts

Composite Parts

Combination of Functions

Favorite Composite Part

9xGSP-GFP(BBa_K3064026)

In order to improve this part, this year we have made a series of modification based on the Minimum TATA-box promoter designed by Daniel Tang of Stanford BIOE44 - S11.(BBa_M50098). Due to the low efficiency of TATA box promoter, we shorten the sequence into only minp. In addition, we also added glucose-sensing fragment to enhance the part’s initiation strength, as well as glucose-sensing function. With GFP, the part’s function can be better detected.(Figure 2)

To conduct the later function test, we set two different groups to conduct the transfection. One is pcDNA3.1-9xGSP-GFP and the other one is plv-mcherry as the internal control. We transfected 300μg into HepG2 cells, which were cultured on 24-hole plate. When 90 percent were mixed, we begin the transfection. At the very first beginning, we starved the HepG2 cells with DMEM for 2 hours before transfection begins.After transfection 12h, we starved the cells with glucose-free culture and stimulate with 20mM glucose concentration culture after 6 more hours. Glucose stimulation intensity was controlled at 20mM. Samples were tested after transfection of 48h.(Figure 1)

Figure 2. The structure diagram of the 9xGSP-GFP part.

Figure 1. Steps of transfection and function test.

Our Composite Parts



Our Composite Parts

NAMETYPEDISCRIPTIONLENGTHSUBMITTED
Part:BBa_K3064005 Composite3XGlucose Sensing Promter702yes
BBa_K3064011 Composite6XGlucose Sensing Promoter345yes
BBa_K3064012 Composite9XGlucose Sensing Promoter1308yes
BBa_K3064014 CompositehsaGlucagon->GGGGS->hsaIgG-Fc->P2A->HA->hsaTrim21519yes
BBa_K3064016 CompositemmuIgG-Fc->HA-Trim21 1062yes
BBa_K3064022 CompositeHA-tag->mmuTrim211653yes
BBa_K3064024 CompositemmuGlucagon-GGGGS-mmulgG-Fc-P2A-HA-mmuTrim21 87yes
Part:BBa_K3064026 Composite9xGSP-GFP87yes