1. At2G37120: A Gene Exploration
An Arabidopsis Journey Through Experimental Questioning
By Jeremy Ziskind

2. What is the structure of AT2G37120?
112011
DNA Pool 5
123257
118631
Expressed in Suspensor of Scarlet Runner Bean
Chromosome 2 of Arabidopsis
-932 base pairs in length
-2 exons
-5’ and 3’ UTRs
What organisms is a potential ortholog found in?
-Oryza sativa, Japanese rice
Madison Insert
SALK Inserts

3. What is the Gene’s Possible Function?
Codes for S1Fa which is most likely a nuclear DNA binding protein
S1Fa important for the binding specificity of a small protein that binds to DNA
Protein only has 70 amino acids (Smallest DNA binding protein ever reported)
Has no homology to any known DNA binding modules
Conclusion: S1Fa belong to a NOVEL CLASS OF DNA BINDING PROTEINS!!!

4. SRB EST vs Arabidopsis
Comparing AT2G37120 gene expression (protein sequence) in Arabidopsis to Scarlet Runner Bean expression
EST: PCSC16872 (42125) Length = 408 Score = 39.6 bits (80), Expect(3) = 2e-09 Identities = 15/21 (71%), Positives = 15/21 (71%) Frame = +2 / -2
Query: 353 MRNVTSRPPITSSTINPGFNP 415
MRNV S PP TS TI PG NP
Sbjct: 146 MRNVNSNPPTTSRTIKPGLNP 84
CONCLUSIONS: SRB EST has 71% region of amino acid homology to Arabidopsis protein sequence

5. Where is S1FA mRNA present in Arabidopsis and SRB?
Conclusions: S1Fa is affected by lec1 because expression patterns are different in the absence of lec1
mRNA accumulation patterns in Scarlet Runner Bean
100 Kb Ladder
PcL1L Control
S1FA Bands
Primer Dimerazation

6. Is there a knockout in Madison Lines? Rounds 1 and 2
Superpool 28
DNA Pool 5
Superpool 28
Control
DNA Pool #5 with a size of about 1400 base pairs
Sequencing results verify that the insert is in DNA Pool #5!
DNA Pool
Screen

7. What is the genotype of Arabidopsis plants with SALK insert #118631?
Negative Water Control
Wild-type Allele Amplification
Mutant Allele Amplification WT
Tubulin
Conclusions: SALK plants 1-9 are homozygous for the wild-type allele and have no T-DNA insertions
Extremely faint Tubulin

8. Further Experiments
Further SALK genotyping must be performed to determine whether At2G37120 is important for seed development
Phenotyping must be done on SALK # plants to determine if the knockouts affect seed development or any other aspect of plant development
DNA Pool #5 must be narrowed down to find the individual line with a knockout in At2G37120