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Genetic Manipulation Techniques in Drosophila for Cellular Analysis

Learn about the experimental setup involving lacZ operon, X-Gal4, UAS-lacZ, and more in Drosophila genetics studies at the University of Manchester. Understand the transformation of imaginal discs into wings and legs through larval preparations embedding and documentation.

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Genetic Manipulation Techniques in Drosophila for Cellular Analysis

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  1. BIOL22332/20972 GENETICS / Dev.Biol RSM MODULE 3 The University of Manchester Faculty of Life Sciences Embedding, documentation & filing

  2. Experimental background What is lacZ? The lac operon of E. coli

  3. β-Gal expressing fly lines X-lacZ enhancer constructs X-Gal4 UAS-lacZ X-lacZ enhancer trap lines

  4. gut proventricle head skeleton on epidermis: wing, haltere, 3rd leg on CNS: 1st & 2nd leg

  5. The transformation of Drosophila imaginal discs into legs and wings

  6. Embedding of larval preparations - transfer larval preparations to a drop of glycerol on a slide - dissect leg and wing discs away and separate on one side of the drop - place 40 μl droplet of glycerol on a fresh (frosted) slide - transfer discs with one shank of the forceps (2-5 wing and 2-5 leg discs) - put on cover slip (see image) - seal all around the cover slip with nail varnish

  7. Filing of slides

  8. 'drawing aid'

  9. Documentation 18.02.2005 John Smith L3, genotype B X-Gal location: B28 arrow: wing margin arrow head: wing hinge

  10. Draw as many of the various genetic strains as possible to have material for the assessed task, i.e. composing a figure and its legend

  11. set to '0' 'A' to 'B' start small focus of condenser use little contrast 'C' to 'D' 'B' to 'C'

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