Tongji iGEM
TongJi iGEM
Results
The outcomes of our experiments.
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Plasmid Construction
We constructed three plasmid successfully: pUAST-pleP-GAL4, pUAST-pleP-GAL80ts and pUAST-UAS-TH.
This is our digest verification map:
This is our digest verification map:
![1% gel electrophoresis of plasmid](https://static.igem.org/mediawiki/2017/e/e3/2017tongji_wiki_image_process_12.png)
pUAST-pleP-GAL4
![1% gel electrophoresis of plasmid](https://static.igem.org/mediawiki/2017/b/b8/2017tongji_wiki_image_process_13.png)
pUAST-pleP-GAL80ts
![1% gel electrophoresis of plasmid](https://static.igem.org/mediawiki/2017/0/0a/2017tongji_wiki_image_process_14.png)
pUAST-UAS-TH
Behavior Tests
Experiment 1: Use real-time PCR to determine whether the expression of TH is higher at 29°C
Figure 1 - The relative expression of TH in modified fruit flies increased significantly.
Experiment 2: Detect male-male courtship when raising the temperature
Mating index (the relative time that the fruit fly use for mating):
Figure 2 - The mating index of the treated group rises significantly in at 29°C.
[time=5minutes, n=5, P<0.01]
Experiment 3: Test the modified Drosophila’s gender preferences
Comparison of time that the fruit flies use for mating male or female flies:
Figure 3 - The modified fruit flies had no preference for males and females when mating.
[time=10minutes, n=8, P>0.5]
Experiment 4: Test whether the modified fruit flies increase their appeal for males
The time that the fruit fly use for mating in five minutes:
There was no courtship in all groups.
Experiment 5: Does the co-culture of wild-type and modified fruit flies influence their courtship with wild-type females?
Comparison of time that the fruit flies use for mating male or female flies:
Figure 4 - The co-culture of wild-type and modified fruit flies did not affect the courtship of wild-type flies.
[time=5minutes, n=5, P>0.5]
Experiment 6: Is the viability of modified fruit flies affected?
The number of live flies in the absence of food and water:
Figure 5 - The survival curve of fruit flies in the absence of food and water
The number of live flies in high temperatures (37°C):
Figure 6 - The survival curve of fruit flies under high temperature
[T=37°C]
The seismo-tube experiment:
Figure 7 - The modified fruit flies crawl higher in the seismo-tube experiment
Experiment 7: The reproductive curve of the modified fruit fly.
Figure 8 - The reproduction curve of the modified fruit fly
[n=5]
Experiment 8: Does the proportion of modified flies have an impact on mating rates of population?
Figure 9 - The impact that proportion of modified flies have on mating rates of population
[n=5]
![TH expression](https://static.igem.org/mediawiki/2017/0/08/2017tongji_wiki_image_result1.png)
Experiment 2: Detect male-male courtship when raising the temperature
Mating index (the relative time that the fruit fly use for mating):
![Mating Index](https://static.igem.org/mediawiki/2017/e/e4/2017tongji_wiki_image_result2.png)
[time=5minutes, n=5, P<0.01]
Experiment 3: Test the modified Drosophila’s gender preferences
Comparison of time that the fruit flies use for mating male or female flies:
![male-male and male-female mating](https://static.igem.org/mediawiki/2017/4/46/2017tongji_wiki_image_result3.png)
[time=10minutes, n=8, P>0.5]
Experiment 4: Test whether the modified fruit flies increase their appeal for males
The time that the fruit fly use for mating in five minutes:
There was no courtship in all groups.
[time=5minutes, n=5]
Experiment 5: Does the co-culture of wild-type and modified fruit flies influence their courtship with wild-type females?
Comparison of time that the fruit flies use for mating male or female flies:
![male-male and male-female mating](https://static.igem.org/mediawiki/2017/7/76/2017tongji_wiki_image_result4.png)
[time=5minutes, n=5, P>0.5]
Experiment 6: Is the viability of modified fruit flies affected?
The number of live flies in the absence of food and water:
![survival without food or water](https://static.igem.org/mediawiki/2017/e/e7/2017tongji_wiki_image_result5.png)
![survival in high temperature](https://static.igem.org/mediawiki/2017/c/ce/2017tongji_wiki_image_result6.png)
[T=37°C]
![survival without food or water](https://static.igem.org/mediawiki/2017/5/55/2017tongji_wiki_image_result7.png)
Experiment 7: The reproductive curve of the modified fruit fly.
![male-male and male-female mating](https://static.igem.org/mediawiki/2017/c/c4/2017tongji_wiki_image_result8.png)
[n=5]
Experiment 8: Does the proportion of modified flies have an impact on mating rates of population?
![male-male and male-female mating](https://static.igem.org/mediawiki/2017/4/4f/2017tongji_wiki_image_result9.png)
[n=5]