Model Organism for Cell division Eukaryote Similar to higher organisms Early embryo Size Transparent Stereotype cell division

First Two rounds of cell Division in Wild Type C. Elegans

Precise Cell Division Timing Cell cycle Early embryogenesis Genes required for DNA replication DNA polymerase machinery Activation of DNA replication Monitor of DNA replication DNA Synthesis

Goal of the project Identify genes that are involved in DNA replication during the cell cycle Approach- utilize cell division mutants Execute DNA replication Activate DNA replication

Temperature Sensitive, Embryonic Lethal Mutant Functional gene Non-functional gene

Cell progression in early cell division (div mutant /P1 delay) Sandra Encalada, 2000 Wild TypeEu2100 or1232

P1 Delay Sandra Encalada, 2000

~ 800 TS-EL 13 confirmed P1 delay 25 candidates 5 strains

or493 or1232 or1332or1579 or1295

The Process in Identifying the Gene Phenotype strain Outcross mutant to wild type Outcross mutant to polymorphic strain for mapping and gene identity

Present Work Identify strain that are P1 delay Phenotype the strain Nomarski DIC 3-5 movies

Results 2 strains are mildly P1 delay or1332 or strains are P1 delay or1232 or strain is mitotic spindle defective (spd) or493

P1 Delay or 1232 or 1579 WT (N2) 0’’ 24.1’ 50.8’ 29.6’ 38.2’ 41.4’34.4’

Mitotic spindle defective (spd) WT (N2) 14.2’ 0’ 29.6’ or ’17.3’33.4’ 0’11.9’ 14.2’

The Process in Identifying the Gene Phenotype strain Outcross mutant to wild type Outcross mutant to polymorphic strain for mapping and gene identity

Present Work and Future Goals Phenotype strain Outcross strain to WT WT F2 Lin-2 or1232 Lin-2 F1 Lin-2 or1232 Lin-2 or1232 Lin-2or1232 Lin-2 or1232

Future direction Phenotype strain Outcross mutant to wild type Outcross mutant to polymorphic strain for mapping and gene identity

Summary Phenotype 3-5 DIC movies of each strain P1 delay: or1232 and or strains were mildly delayed 1 strain was found to be mitotic spindle defective The strains confirmed to be P1, out-crossing to wild type is carried out. In the future, desired outcross mutant strain will be outcross to polymorphic strain for WGS.

Acknowledgements Bruce Bowerman –-PI Joshua Lowry –Mentor Peter, Adam, SPURmies Reza--Post-Doc Chien Hui–- Lab Manager Amy--Graduate student Sean– Post Doc Ava Sophie Diana NSF REU BIO THE Undergraduates

Thank you.