Part:BBa_K1061001

apoptin

Apoptin is a protein that is originally isolated from the chicken anemia virus(CAV). It has been found that expressing this gene can induce apoptosis in a broad spectrum of cancer cells but not in normal cells. Hence it is a very powerful tool in treating cancer. Fusion the TAT protein with apoptin can even achieve by-stander effect, and we try to use it in the most simple device of our project. We have successfully expressed it in Hep G2 cell lines and observe a brilliant apoptosis effect.[1]

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 172
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

calibration

Pre-experiment that we done

apoptosis effect

(Apoptosis phenotype of Apoptin(VP3). The protein was expressed in fusion with a GFP. Compared with 0 hr , apoptosis rose 48 hr after transfection, and GFP vision indicated the transfection efficiency)However, due to technical problem in cell culture,the transfection effciency can not reach such a high level in following experiment.The phenomena become not so obvious,but still exist.To observe that, we must carefully count the cells. So, finally we decided to use the Flow cytometry technique to confirm the apoptosis effect.Details will be show in our wiki.http://2013.igem.org/Team:SYSU-China/Project/Results

Kill range

For each suicide gene, it is important to know which cancer cells and normal cells it will kill.So that people can speficially choose gene that will be functional in their project when duelling about cancer. And we define this new feature for each suicide gene that we submitted. The list will be changed due to more and more experiment will be done. We highly recommend people to adopt this characteristic as important characteristic of suicide gene.

Functional Parameters: Austin_UTexas

BBa_K1061001 parameters

Burden Imposed by this Part:

Burden is the percent reduction in the growth rate of E. coli cells transformed with a plasmid containing this BioBrick (± values are 95% confidence limits). This BioBrick did not exhibit a burden that was significantly greater than zero (i.e., it appears to have little to no impact on growth). Therefore, users can depend on this part to remain stable for many bacterial cell divisions and in large culture volumes. Refer to any one of the BBa_K3174002 - BBa_K3174007 pages for more information on the methods, an explanation of the sources of burden, and other conclusions from a large-scale measurement project conducted by the 2019 Austin_UTexas team.

This functional parameter was added by the 2020 Austin_UTexas team.

References:

[1] Claude Backendorf et al,Apoptin: Therapeutic Potential of an Early Sensor of Carcinogenic Transformation,Annu. Rev. Pharmacol. Toxicol. 2008. 48:143–69