Difference between revisions of "Part:BBa K1061001"
Line 10: Line 10:
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1061001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1061001 SequenceAndFeatures</partinfo>
−
 
+
====Calibration====
 +
Pre-experiment that we done
 +
[[Image:Suicidegene-vp3.png|thumb|center|500px|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.Detail will be show in our wiki.http://2013.igem.org/Team:SYSU-China/Project/Results
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 16:58, 26 September 2013

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.Detail will be show in our wiki.http://2013.igem.org/Team:SYSU-China/Project/Results

References:

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