Part:BBa_K758006

UAS EGFP, this part expresses EGFP in the presence of GAL4 protein.

This part consists of five UAS sequences, EGFP, SV40 polyadenylation signal. This part is similary to BBa_K758005 because this part and BBa_K758005 altered LacZ to EGFP are almost the same. So, if GAL4 exists appropriately, this part expresses EGFP.

Experiments and Results

We transfected this plasmid to Drosophila culture cells under two different conditions. We also transfected pAct5C-GAL4 plasmids which express GAL4 proteins in a culture cell line but not in other cell line.
If the BBa_K758006 was assembled accurately, this plasmid expresses enough EGFP by activation of GAL4 protein. So the cell line that were co-transfected with pAct5C-GAL4 shows strong fluorescence, than the one without co-transfection with pAct5C-GAL4.

Then we conducted experiment mentioned above. We made two groups of Drosophila culture cells. One group had been transfected with BBa_K758006 and pAct5C-GAL4, and the other one had been transfected with BBa_K758006 alone. And we calculated ratios between the number of green lighted cells to total cells about both groups.

As results shown below, some green cells were detected among the cells co-transfected with both plasmids. But few green cells were detected among the cells transfected BBa_K758006 alone.
And we also compiled statistics on the ratio of green lighted cells. In consequence, in the group transfected with pAct5C-GAL4, there were 45 green colored cells (12.6%) among 356 cells in total. And, without pAct5C-GAL4, there were only 2 cells were colored green (it is 0.5%) among 350 cells in total.

These results indicate that BBa_K758006 was activated by GAL4 protein and BB_K758006 EGFP expression was induced by GAL4 protein.
In these circumstances, BBa_K758006 was working as expected.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 1130
  Illegal PstI site found at 4
  Illegal PstI site found at 247
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 4
  Illegal PstI site found at 247
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 377
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 1130
  Illegal PstI site found at 4
  Illegal PstI site found at 247
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 1130
  Illegal PstI site found at 4
  Illegal PstI site found at 247
  
• 1000
  COMPATIBLE WITH RFC[1000]