Upload
kathryn-spencer
View
224
Download
0
Tags:
Embed Size (px)
Citation preview
A Hypothesis for the function of gene AT4G23180 in A.
thalianaBy Nicole Foxworth and Deborah
Lee(Ether Fowl Ox)
CREATING A GENE MODEL
Lab 2.2
Predicted Gene Structure
GENE KNOCKOUT MUTATION
Lab 3.1
SALK 116653 Will Effectively Knock Out Our Gene
SALK 116653 is within an exon and will likely cause a knockout mutation of At4g23180.
1860bp1395bp
Key
T-DNA Insertion Site
SALK 116653
R L
MUTANT PLANT OBSERVATIONS
Lab 3.2
Clear Difference in Growth and Development
SALK Plants
Day 7- Hypocotylpresent
Day 15- 4 true leaves
Wild Type Plants
Day 7- Cotyledons &2 true leaves emerging
Day 15- 5 true leaves &2 more emerging
SALK Plants HaveSlower Onset of Leaf
Development
Effect of Low Fe Medium is Inconclusive
Low Fe Medium
Day 9
- No seeds germinated
MS Medium
Day 9
- One SALK plant germinated
RESEARCHING GENE FUNCTION: BIOINFORMATICS
Lab 4.1
Protein Kinase Catalytic Domain is a Conserved
Domain in our Gene
CRK10/RLK4 is a Perfect Match to our Predicted Protein Sequence
RESEARCHING GENE FUNCTION: LITERATURE SEARCH
Lab 4.2
RLK4 is Expressed in the Shoot Apical Meristem and
Lateral Root• Review paper entitled
“Receptor Kinase Signaling in Plant Development” (Becraft 2002)
• Overview of many proteins in the RLK family, but does not indicate a specific function for RLK4
CRK10 Plays a Role in Plant Defense
• Chen et. al. explored entire CRK family–Found that many CRKs are involved
in plant defense responses–CRK10 is not soley responsible for
plant defense, but plays a role. • Could be due to functional redundancy
of closely related genes
RLK4 is Involved in Response to Pathogen Infection
• Du and Chen find evidence that RLK4 is highly expressed in leaves after pathogen infection.
MICROARRAY RESOURCES
Lab 5.1-5.2
Location of At4g23180 Expression
1st Experiment Idea: Seedling Root Response to Salinity
Stress
Focused Experiment on the Effects of Wounding
Gene At4g23180 expression increases in response to leaf wounding/aphid feeding.
HYPOTHESIS TESTINGLab 5.4-5.6
Hypothesis of Gene Function: Plant Defense
• We hypothesize that our gene is involved in plant defensive response to wounding and is expressed in the leaves.
• We designed an experiment to test this hypothesis.– Poked holes in plant leaves and tested
level of gene expression after 30 minutes.
Experiment Yielded Inconclusive Results
A D
B C
Future Directions
• We maintain our hypothesis that our gene is involved in plant defensive response
• Repeat our designed experiment with the following changes:– Bigger sample size– Quality assessment of RNA extracted
through gel electrophoresis– Design new primers within exons that flank
an intron but are not split in half by one
References
• Becraft, Philip W. "Receptor Kinase Signaling In Plant Development." Annual Review of Cell and Developmental Biology 18.1 (2002): 163-92. Print.
• Chen, Kegui, Baofang Fan, Liqun Du, and Zhixiang Chen. "Activation of Hypersensitive Cell Death by Pathogen-induced Receptor-like Protein Kinases from Arabidopsis." Plant Molecular Biology 56.2 (2004): 271-83. Print.
• Du, Liqun, and Zhixiang Chen. "Identification of Genes Encoding Receptor-like Protein Kinases as Possible Targets of Pathogen- and Salicylic Acid-induced WRKY DNA-binding Proteins in Arabidopsis." The Plant Journal 24.6 (2000): 837-47. Print.