BIO205

Weinberg’s Lab,”used 3T3 mouse cell lineswas first looking for cells that lose contact inhibition (focus) that piled on top of each other (most didn’t)he caused a tumorthen isolated DNA from that tumor, extracted it, then cloned DNA that’s in there. 99.99% of sequence will be mouse, but 1 gene is human. Can identify human DNA and distinguish from mouse DNA by molecular marker (weinberg able to take DNA isolated from human carcinoma cell–>this is how Ras^D first identified oncogene found in this cancer line and was able to transfer 3T3 cells and make them tumoregenic)Could have also changed the selection you use. original carcinoma isolating RasD, losing contact inhibition was first they looked for…can go back to same tumor DNA, keep transfection, instead of losing contact inhibition

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Multi-Hit Model,”Cancer is a disease of aging. It’s not a disease that affects the young people (rare)bottom graph: how many days during the life of the mouse it’ll go through while tumor freeMyc mice relatively tumor free after 120 days, then gradual decline in # of tumor free individualsCompared to Ras, finding that mice that harbor Ras oncogene start acquiring tumors much, much earlierIf cross two strains, now have mice that have both oncogenes. Ras operating much further upstream than Myc. When put together, get synergistic effect, causing rapid onset of tumorigenesis

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“You’re performing the Ames test on two known carcinogens; Benzo(a)pyrene (BaP) and methyl nitrosourea (MNU). If you treat His-Salmonella with MNU, you generate many His+ colonies. In contrast, if you treat His auxotrophic Salmonella with BaP, you generate no His prototrophs. However, if you first incubate the BaP with a liver extract before adding it to the His auxotroph Salmonella, you generate many His prototrophs. How do BaP and MNU differ?”

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