Studies of Wnt Signalling in Drosophila:
Subtitle: Screens for Candidate Transcripts of a New Gene Involved in Transduction and Genetic and Molecular Analysis of a Potential Target
Publisher: University of Antwerp
Published: 1999
Place of publication: Antwerp, Belgium
Language: eng
Format: Paperback
Number of pages: 84
Description: MSc - 1999
Table of contents: label: Dedication; label: Table of Contents; title: 1
label: Abbreviations; title: 2
label: Nederlandse Samenvatting; title: 6
label: English Summary; title: 7
label: Introduction; title: 7
label: I. Peripheral sensory development; title: 7
label: 1. Introduction; title: 8
label: 2. The genetic mechanisms underlying bristle patterns; title: 8
label: A. Overview; title: 10
label: B. Expression of achaete and scute in the proneural clusters; title: 10
label: C. Selection of the SMC in the proneural cluster; title: 12
label: D. Asymmetric division of the SMC; title: 12
label: E. Development of the proneural clusters relies on a prepattern; title: 13
label: II. Wingless in Drosophila development; title: 15
label: III. Wnt signal transduction pathway; title: 15
label: 1. Wg/Wnt-1 pathway; title: 17
label: 2. Conservation of the Wg/Wnt-1 pathway; title: 18
label: IV. Wnts in vertebrate development; title: 19
label: V. Wnts in human disease; title: 19
label: 1. Wnt signalling and colon cancer; title: 20
label: 2. Wnt signalling and Alzheimer’s Disease; title: 21
label: 3. GSK-3 and diabetes; title: 22
label: VI. Objectives; title: 24
label: Materials and Methods; title: 24
label: I. Kiwi; title: 24
label: 1. General; title: 24
label: A. Preparing ampicillin (kanamycin) solutions; title: 24
label: B. Pouring plates; title: 24
label: C. Overnights; title: 25
label: D. Small-scale preparations of plasmid DNA; title: 25
label: E. Glycerol stocks; title: 26
label: F. Competent cells and transformation; title: 26
label: a. Competent cells; title: 26
label: b. Transformations; title: 26
label: G. Electrophoresis; title: 27
label: H. Fixing DNA to membranes; title: 27
label: I. Hybridisation; title: 27
label: a. Introduction; title: 28
label: b. Prehybridisation; title: 28
label: c. Preparing radioactive probe; title: 28
label: d. Assay the radioactive incorporation; title: 29
label: e. Hybridisation; title: 29
label: f. Washes; title: 29
label: g. Mounting of membrane; title: 30
label: 2. Screening a library; title: 31
label: 3. Tn1000 approach; title: 31
label: A. Mating; title: 32
label: B. Identification of inserts; title: 32
label: a. Overnights; title: 32
label: b. Cosmid preps; title: 33
label: c. Digest; title: 33
label: d. Primer synthesis; title: 34
label: e. Annealing of the vectorette; title: 35
label: f. Ligation; title: 35
label: g. PCR; title: 38
label: h. Running a gel; title: 38
label: i. The blotting conditions; title: 38
label: i1. Type of blotting procedure; title: 39
label: i2. Denaturation step and blotting buffer; title: 39
label: i3. Brand of membrane; title: 39
label: i4. Optimised blotting protocol; title: 40
label: II. ED17; title: 40
label: 1. Plasmid rescue; title: 40
label: A. Isolation of DNA from adult flies; title: 41
label: B. Plasmid rescue; title: 41
label: 2. Fly crosses; title: 43
label: 3. Staining; title: 43
label: A. X-gal staining and light microscopy; title: 44
label: B. Ab-staining and epifluorescence/confocal microscopy; title: 45
label: Results; title: 45
label: I. Kiwi; title: 45
label: 1. Introduction; title: 45
label: 2. Mapping of kiwi; title: 45
label: A. Deletion mapping; title: 47
label: B. Inversion; title: 47
label: C. Contigs; title: 48
label: 3. Screening of a cDNA library from 0h - 4h embryos; title: 50
label: 4. Tn1000 approach; title: 55
label: II. ED17; title: 55
label: 1. Enhancer trapping; title: 55
label: 2. X-gal and antibody staining of larval imaginal wing discs; title: 58
label: 3. Excision lines; title: 60
label: Discussion; title: 60
label: I. The kiwi locus; title: 60
label: 1. Improving the transposon approach; title: 60
label: 2. Alternative approaches to retrieve coding region; title: 61
label: 3. Information from in vitro transcribed RNA; title: 62
label: II. The ED17 locus; title: 62
label: 1. Study of the phenotype of excision lines of P[ED17]; title: 62
label: 2. Mapping of the P[ED17] insertion site; title: 63
label: 3. Genetic model explaining the ED17 expression pattern; title: 66
label: Acknowledgements; title: 67
label: References; title: 71
label: Appendix A; title: 73
label: Appendix B; title: 78
label: Appendix C