Transgenic Fly Biology Lab, answer 8 questions in the attached file.

transgenicflylab.pptx

Transgenic fly lab

Two parts

Week 1 – make a transgenic fly

Week 2 – Use transgenic flies to study circadian rhythms and genetics

What’s the point? – to use luciferase expression to see when and where the period gene is expressed in fruit flies.

Click here to learn about the plasmid(s)

Look under Notebook of Part 1

Step 1: Preparing the DNA - transposons

Insert plasmid – already been constructed for you

Scientist “high-jacked” transposable elements

Transposons – Can move around within a genome

The P element is the transposable element of the transposon

Transposase – enzyme that actually cuts out the P element (and surrounding genes).

Step 1: Preparing the DNA – the plasmid

P element – allow region to jump into the fly genome

Per – period gene promoter

Luc – luciferase gene

Mini-white – promoter and coding region

Changes the eye color in offspring to red color (look WT)

Step 1: Preparing the DNA – second plasmid

Transposase DNA on a second plasmid

You need both to get Construct DNA into the fly genome

Transposase DNA plasmid will not insert into the genome

Click here to learn about the plasmid sequence of per promoter

Look under Notebook of Part 1

Complete the

“A. Confirm that the construct

DNA sequence is from the per gene.”

Step 5: Selecting embryos

We inject embryos with both plasmids

Will the injected embryos have red eyes or WT?

Will the offspring of the embryos injected have red eyes or white eyes?

Week 2: Use Transgenic Flies

Pick the red eye flies and use these to test for period gene expression.