Favorite Composite Part
9xGSP-GFP
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
| NAME | TYPE | DISCRIPTION | LENGTH | SUBMITTED |
| Part:BBa_K3064005 | Composite | 3XGlucose Sensing Promter | 702 | yes |
| BBa_K3064011 | Composite | 6XGlucose Sensing Promoter | 345 | yes |
| BBa_K3064012 | Composite | 9XGlucose Sensing Promoter | 1308 | yes |
| BBa_K3064014 | Composite | hsaGlucagon->GGGGS->hsaIgG-Fc->P2A->HA->hsaTrim21 | 519 | yes |
| BBa_K3064016 | Composite | mmuIgG-Fc->HA-Trim21 | 1062 | yes |
| BBa_K3064022 | Composite | HA-tag->mmuTrim21 | 1653 | yes |
| BBa_K3064024 | Composite | mmuGlucagon-GGGGS-mmulgG-Fc-P2A-HA-mmuTrim21 | 87 | yes |
| Part:BBa_K3064026 | Composite | 9xGSP-GFP | 87 | yes |