BIOL3108 Lecture Notes - Lecture 6: 3T3 Cells, Transfection, Southern Blot
Document Summary
This is just a movement away from the idea that cancer can only be caused by viruses - carcinogens can cause cancer. Free radicals - often form due to radiation, etc. 1972: transfection of mutated dna can transform cells and detect oncogenes. Method: dna is extracted and co-precipitated to generate calcium phosphate crystals that are added to a monolayer of normal cells. If a transforming gene (oncogene) is present in the donor dna and is incorporated into the genome to transform a recipient cell this will over-proliferate to form a clump (colony) Injection of these cells into a host mouse and resulting tumour formation can be used to confirm the transformed state. Conclusion: exposure of normal mouse cells to 3-mc carcinogen caused a single mutant oncogenic allele and this was sufficient to transform recipient nih 3t3 cells. Transfection experiments provided evidence that oncogenes can arise in the genomes of cells through non- viral mechanisms.