1. 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 blade
Leaf sheath

2. 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

4. Experimental Design One hour BrdU Pulse Fix Cells and Isolate Nuclei
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

5. Arabidopsis Chromosome 4
genes
transposable elements
nucleolar
organizing region
centromere

6. Replication Timing Profile
log (early/late) L
chromosome coordinate

7. Origin assembly Pre-replication complex licensing
Initiation complexes assemble in late M and early G1
ORC - origin recognition complex
MCM - replicative DNA helicase
CDT1/CDC6 - MCM loader
licensing

8. MCM5/7 are released during mitosis
Merge
DAPI
DIC
MCM5
MCM7
N. tabacum cultured cells (Nt-1)

9. How is licensing regulated in plants?X
CDT1
Proteosome
CDKA P
A different CDT regulatory protein?
Geminin
ORC1 regulation?
MCM subunits X
Other mechanisms?
cytoplasm
nucleus

10. Exploring Genetics across the Middle School Science and Math Curriculum
Classroom visits

11. Exploring Genetics across the Middle School Science and Math Curriculum
Classroom visits

12. Genetics Kit Activities Heredity & Genetics Scavenger Hunt
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

13. Activity 2: Survey of Inherited Traits
Activity 5: Genetic Offspring Models
Activity 6: Extracting Wheat DNA
Activity 8: Day in the Life of DNA

14. Genetics Independent Study Group