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 in Dubna Dzelepov Laboratory of Nuclear Problems 2014 Student Practice
Our goal: to detect the nature and location of DNA alterations induced by γ-rays at the yellow gene of Drosophila melanogaster
Mutations - somatic / heritable - point / chromosomal substitution insertion deletion - effects of mutations: silent / missense / nonsense
Effects of ionizing radiation on DNA gamma neutrons
Effects of ionizing radiation on DNA
Drosophila melanogaster - model organism - short life cycle (10 days) - only 4 chromosomes - genom sequenced - giant chromosomes
brown eyes cinnabar eyes black vermilion eyes wild type white eyes yellow
yellow gene - protein coding gene - 4.7 kb - paratelomeric
Conditions of irradiation radiation: 60 Co gamma-rays source: “Gamma-cell-220” plate dose: 40 Gy dose rate: 5,7 Gy/min linear energy transfer: 0.3 keV/micron
How to detect changes in DNA? DNA isolation amplification of a DNA fragment (PCR) electrophoresis
DNA isolation - cell lysis - DNA sorption on silica particles - DNA purification - DNA extraction (from silica particles)
PCR
PCR Electrophoresis
Results
Some more results
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?)
Thank you for your attention!
Recommend
More recommend
