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PCR-assay of the mutational lesions induced by ionizing radiation at - - PowerPoint PPT Presentation
PCR-assay of the mutational lesions induced by ionizing radiation at - - PowerPoint PPT Presentation
PCR-assay of the mutational lesions induced by ionizing radiation at the paratelomeric yellow gene of Drosophila melanogaster Martina Luzova Kaja Cegielka supervisors: Igor Alexandrov Kristina Afanasayeva Joint Institute for Nuclear Research
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Mutations
- somatic / heritable
- point / chromosomal
substitution insertion deletion
- effects of mutations:
silent / missense / nonsense
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Effects of ionizing radiation on DNA
neutrons gamma
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Effects of ionizing radiation on DNA
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Drosophila melanogaster
- model organism
- short life cycle (10 days)
- only 4 chromosomes
- genom sequenced
- giant chromosomes
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wild type
yellow
white eyes black brown eyes cinnabar eyes vermilion eyes
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yellow gene
- protein coding gene
- 4.7 kb
- paratelomeric
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Conditions of irradiation
radiation: 60Co gamma-rays source: “Gamma-cell-220” plate dose: 40 Gy dose rate: 5,7 Gy/min linear energy transfer: 0.3 keV/micron
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How to detect changes in DNA?
DNA isolation amplification of a DNA fragment (PCR) electrophoresis
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DNA isolation
- cell lysis
- DNA sorption on silica particles
- DNA purification
- DNA extraction (from silica particles)
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PCR
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Electrophoresis PCR
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Results
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Some more results
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Results - summary
- no big deletions of the yellow gene
- no chromosome breaks inside the gene
- point mutations in y94 and y25
- further studies necessary (sequencing?)
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