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Why study DNA replication?. Attractive model for multicellular eukaryotes Small, sequenced genomes Genomic tools available Rapidly dividing cell cultures Tractable developmental programs. Arabidopsis. Rice. Intercalary meristem. Leaf sheath. Leaf blade. Why study DNA replication?.
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Why study DNA replication? • Attractive model for multicellular eukaryotes • Small, sequenced genomes • Genomic tools available • Rapidly dividing cell cultures • Tractable developmental programs Arabidopsis Rice Intercalary meristem Leaf sheath Leaf blade
Why study DNA replication? • Potential practical applications • Genetic engineering of artificial chromosomes • Manipulating the endocycle • New strategies for geminivirus resistance Cotton leaf curl disease Tomato leaf curl disease
One hour BrdU Pulse Fix Cells and Isolate Nuclei Experimental Design FACS “synchronization” IP BrdU-labeled DNA Label IP and Input DNA with Cy3/5 Hybridize to Chr4 Tiling Array 20000 probes 1 kb PCR products 95% coverage Normalize Smooth
Arabidopsis Chromosome 4 genes transposable elements nucleolar organizing region centromere
L E L E L E L E L Replication Timing Profile E log (early/late) L chromosome coordinate
Origin assembly Pre-replication complex Initiation complexes assemble in late M and early G1 ORC - origin recognition complex MCM - replicative DNA helicase CDT1/CDC6 - MCM loader licensing
MCM5/7 are released during mitosis DAPI DIC Merge MCM5 MCM7 N. tabacumcultured cells (Nt-1)
Proteosome CDKA P ORC1 regulation? CDT1 MCM subunits How is licensing regulated in plants? ? X A different CDT regulatory protein? Geminin X Other mechanisms? cytoplasm nucleus
Exploring Genetics across the Middle School Science and Math Curriculum Classroom visits
Exploring Genetics across the Middle School Science and Math Curriculum Classroom visits
Dr. Jeff Batten - Overview science activities Genetics Kit Activities Heredity & Genetics Scavenger Hunt Personal Survey of Inherited Human Traits Modeling Mendel Punnett Squares Genetic Offspring Models Extracting DNA from Wheat Germ DNA, Genes, and Proteins (Sickle Cell Disease) A Day in the Life of DNA Genetic Disorders Genetic vs. Environmental Influences on Traits
Activity 2: Survey of Inherited Traits Activity 5: Genetic Offspring Models Activity 6: Extracting Wheat DNA Activity 8: Day in the Life of DNA