Transgenic Fly Biology Lab,
View This
Promega video about reporter genes assays:
https://www.youtube.com/watch?v=xqTudg1WdmY
Youtube video about how to make transgenic flies:
https://www.youtube.com/watch?v=1CJfj1GG_f8
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Transgenic organisms are defined as a genetically modified organisms that contains DNA from two different genomes. A transgenic organism (or knock-in organism) contains a gene which is overexpressed for the purpose of ascertaining the gene function and location. The method of creating a transgenic organism is the same as creating a knock-out organism. A plasmid vector must be used for inserting the gene of interest into the genome. The vector is introduced into the gametic cells of the organism and the experssion is confirmed in the offspring.
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"Make Transgenic Flies" will be completed first then do the "Use Transgenic Flies".
Copy/past this link into your browser. Click on "Launch Interactive." Complete the lab and answer the questions on the worksheet. Remember to read all the information.
https://www.biointeractive.org/classroom-resources/transgenic-fly-virtual-lab
Make Transgenic fly lab 1. (1 pt) You must submit a screen shot of your successful mutant flies at the end of the “Make Transgenic fly lab”. This screenshot will be submitted to a Discussion after completing the lab. When submitting the worksheet – please answer: Where you successful in your experiment? What went wrong? 2. (0.5 pts) What gene are you attaching to the period gene to visualize when transcription occurs? Which organism does this gene come from? 3. (0.5 pts) What key molecular “tool” is used to create the construct of per/luc genes that will jump into the Drosophila genome? 4. (0.5 pts) How will we know if the construct has incorporated into the Drosophila genome? 5. (1 pt) Complete part A of Performing a sequence analysis in the “Make Transgeneic lab” to confirm that the correct period gene is incorporated into the plasmid construct. In the BLAST sequence, what color means a good match? What chromosome is period gene found in Drosophila? Testing Transgenic flies 1. (0.5 pts) About what time of the day does the fly have peak luciferase expression? 2. (0.5 pts) Does the Per gene continue to oscillate regardless of external light/dark cues? 3. (0.5 pts) What time did the peak luciferase expression become after the shift in light/dark cycle?