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Establishment of CAG-GFP-LSL-Cdc6OE-LacZ Mouse Model and Mutant Kras Activation

This study focuses on establishing a transgenic mouse model with whole-body ectopic expression of Cdc6 and LacZ using synthetic promoters. The model involves Cre activation, mutant Kras activation, and PCR detection in newborn mice.

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Establishment of CAG-GFP-LSL-Cdc6OE-LacZ Mouse Model and Mutant Kras Activation

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  1. CAG-GFP-LSL-Cdc6OE-LacZ cdc6b LacZ CAG: Synthetic promoter (CMV early enhancer/chicken β actin) LSL: Lox-Stop-Lox

  2. Cdc6 whole-body or tissue-specific ectopic expression loxP loxP CAG eGFP STOP cdc6b IRES Β-gal loxP loxP CAG eGFP STOP cdc6b IRES Β-gal Whole-body or tissue-specific Cre activation No Cre expression CAG eGFP CAG cdc6b IRES Β-gal GFP expression in all tissues Β-gal cdc6b Cdc6b and β-galactosidase expression in Cre activated cells CAG eGFP GFP expression in non-Cre activated cells

  3. Establishment of CAG-GFP-LSL-Cdc6OE-LacZ mouse Cdc6 transgene Foster mother Two pronuclei Founder of the Cdc6 mouse strain > Cdc6 +/- > > >50% BL6 > Cdc6 +/- > We are at this stage > >75% BL6 Cdc6 +/- approx 100% BL6

  4. Cdc6 ectopic expression and mutant Kras activation in the whole body CAG-GFP-LSL-Cdc6OE-LacZ CAG-CreERTM LSL-KrasG12D CAG-GFP-LSL-cdc6OE-LacZ; CAG-CreERTM + Tamoxifen + Tamoxifen Cre expression Cre expression Cdc6 and β-Galactosidase expression Mutant KrasG12D, Cdc6 and β-Galactosidase expression

  5. PCR to detect transgenic mice Newborn mice L 1 2 3 4 5 6 P L: Ladder P: Plasmid, positive control 1: Transgenic mouse 2-6: Non-transgenic mice

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