Genome 371 Lecture 8 - Genome 371 Lecture 8 1/26/11 4:16 PM...

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1/26/11 4:16 PM Genome 371 Lecture 8 Page 1 of 11 http://courses.washington.edu/au371mkr/resources/lecture_notes/lecture_08.html Lecture 8 Analyzing mutants 25 Oct 2010 Lecture 8 handout (pdf) Subscribe to podcast: Slide 1 Slide 2 Slide 3 The fact that the two elements of a transposon can be separated lets us control and use transposons as a mutagenic tool, as we'll see next. Home Course mechanics Help hours Calendar Syllabus Lectures, Podcasts Quiz Sections Practice problems Exams GoPost Send email to class Useful links The Gradiator
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1/26/11 4:16 PM Genome 371 Lecture 8 Page 2 of 11 http://courses.washington.edu/au371mkr/resources/lecture_notes/lecture_08.html Slide 4 Slide 5 Slide 6 Reminder: The goal is to make mutations that potentially could disrupt the function of some process of interest. The hope is that when transposition happens, the transposon will jump into or next to some gene and disrupt its function. For example: if we are interested in understanding how fruit flies smell bananas, we could look for mutants that no longer are able to smell bananas. Since the flies were able to smell bananas before we triggered the mutation and the mutant progeny are unable to smell bananas, we can conclude that the transposon disrupted some gene that is needed for banana-detection. Slide 7
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1/26/11 4:16 PM Genome 371 Lecture 8 Page 3 of 11 http://courses.washington.edu/au371mkr/resources/lecture_notes/lecture_08.html Slide 8 Slide 9 Slide 10 Slide 11 To have control over transposition (i.e., in which animals it happens), geneticists modified a Drosophila transposon called the "P element" to suit their needs. The transposon was separated into its two critical components. One resulting version, called the Jump-starter, has its transposase gene intact so it can still encode active transposase enzyme -- but one of the inverted repeats was deleted. Therefore, although this crippled " Jump-starter " transposon can make transposase, it cannot itself move ("jump"). In the other version, called P[lacZ,w + ] or just P[w + ] , the inverted repeats were left untouched and intact, but the transposase gene was removed and replaced by a wild type copy of the Drosophila white gene ( w + ) and a couple of other sequences we'll see again later. Because it lacks a transposase, this version is also crippled - - it cannot make itself jump. .. but if transposase enzyme is provided from elsewhere, that transposase can act on the intact inverted repeats and initiate a jump.
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1/26/11 4:16 PM Genome 371 Lecture 8 Page 4 of 11 http://courses.washington.edu/au371mkr/resources/lecture_notes/lecture_08.html Slide 12 See practice question on p.6-135. Slide 13
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Genome 371 Lecture 8 - Genome 371 Lecture 8 1/26/11 4:16 PM...

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