Arabidopsis thealiana AT3G56230 AT4G18650
description
Transcript of Arabidopsis thealiana AT3G56230 AT4G18650
Arabidopsis thealianaAT3G56230AT4G18650
What are the functions of them?
Are they important for seed development?
Spring 2004 Mariko Onozawa
What is AT3G56230?
・ About 1200bps.
・ 5’UTR.
・ Three SALK lines.
・ This protein containing BTB/POZ domain.
What is AT4G18650?
・ About 1400bps.
・ 5’ and 3’UTR.
・ No SALK line.
・ This protein is related to transcription
factor and has weak similarity to
TGAGG-sequence specific DNA-
binding protein(TGA-2.1).
FV RVFV RV
mRNA
*Find the pattern of mRNA concentration
RT-PCR (SRB)
Genechip database
Gene
* Find KOs of my gene
Madison lines
SALK lines
Phenotype
*Study a phenotype of plants which containing the KO
Check genotypes of the plants (SALK)
mRNA Gene Phenotype
Reverse Genetics
RT-PCR result using SRB (AT3G56230)
Genechip data*RT-PCR result says that the gene which is
homologous toAT3G56230 is expressed at all
stages and organs of SRB.
*Genechip data says that AT3G56230
is expressed only at Mature green seed,
Seedling (3DAI), Roots and stem.
0
500
1000
1500
2000
2500
3000
Ovu
les
24-Hr S
eeds
Mid-
Matu
ration
Matu
re G
S
Post-
Matu
ration G
S
seedlin
g (3DAI)
Leave
s
Roots
Ste
m
AT4G18650AT3G56230
A border between presence and absence
Leaves
+R
T
-RT -RT -RT -RT -RT-RT -RT -RTStem
+
RT
Flow
er +
RT
Ovules
+R
T
Em
bryo +
RT
Axes
+R
T
Leaves
+R
T
Stem
+
RT
control
gene specific
Is there any KO of my gene in Madison lines?*AT3G56230
I couldn’t find any KO of AT3G56230.The insertion site is upstream of the gene. (-594bp)
*AT4G18650
I couldn’t find any KO of AT4G18650.The insertion site is downstream of the gene.
FV RV
R19
FV RV
R15
Checking phenotypes of the plants
T-DNA,homo 1,4WT, homo 2,3,5,6
①WT ①T-DNA ②WT, T-DNA
Only tubilan’s bands
nothing
Some samples don’t have any band.
①WT ①T-DNA②WT, T-DNA
Only tubulin’s bandsNothing
Some seedlings don’t have any band.
③WT, T-DNA
④WT, T-DNAsmears
1 2 3 4 5 6WC 1 2 3 4 5 6WC
③WT, T-DNA
smears
④WT T-DNA
1 2 3 4 5 6 WC 1 2 3 4 5 6 WC
I couldn’t find any significant difference between WT and mutant plants.
T-DNA Wild Type
What should be done next to study the functions?
*Get more plants which have a insertion in the genes.*Observe and study the knockout plants.*Study with other knockout lines.
The final goal is to answer these questions.
*What is the function of the protein?*What is the structure of the protein?*How it is regulated?
What we did in this class is only the beginning of the research.