Download Outcross mutant to polymorphic strain for mapping and gene identity

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
Goal of the project
 Identify genes that are involved in DNA replication
during the cell cycle
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Approach- utilize cell division mutants
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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)
Wild Type
Eu2100 or1232
Sandra Encalada, 2000
P1 Delay
Sandra Encalada, 2000
~ 800 TS-EL
13 confirmed P1
delay
25 candidates
5 strains
5 strains
or493
or1295
or1579
or1232
or1332
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
 or1579
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 2 strains are P1 delay
or1232
 or1579

 1 strain is mitotic spindle defective (spd)
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or493
P1 Delay
24.1’
29.6’
0’’
34.4’
41.4’
0’’
38.2’
50.8’
or 1232
or 1579
WT (N2)
0’’
Mitotic spindle defective (spd)
0’
or493
WT (N2)
14.2’
17.3’
14.2’
29.6’
0’
11.9’
21.7’
33.4’
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
Lin-2
or1232
Lin-2
or1232
Lin-2
Outcross strain to WT
WT
or1232
Lin-2
F2
Lin-2
Lin-2
or1232
or1232
or1232
F1
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 or1579
 2 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
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Bruce Bowerman –-PI
Joshua Lowry –Mentor
Peter, Adam, SPURmies
Reza--Post-Doc
Chien Hui–- Lab Manager
Amy--Graduate student
Sean– Post Doc
Ava
THE
Sophie
Undergraduates
Diana
NSF REU BIO 1063144
Thank you.