Part:BBa_K2089002
cop1 promoter from Arabidopsis
Represses photomorphogenesis and induces skotomorphogenesis in the dark. Contains a ring finger zinc-binding motif, a coiled-coil domain, and several WD-40 repeats, similar to G-beta proteins. The C-terminus has homology to TAFII80, a subunit of the TFIID component of the RNA polymerase II of Drosophila. Nuclear localization in the dark and cytoplasmic in the light.
To test the efficiency of this promoter,we construct the recombinant plasmid using this promoter and hhl1(BBa_K2089005) and then we tranfect it into protoplasts.We incubate the protoplasts at 2 groups of light intensities to induce the expression of hhl1,which can show the efficiency of this promoter.
We use semi-quantitive RT-PCR to amplify gene hhl1 from mRNA in protoplasts，the concentration of amplified gene hhl1 of each group is measured by ultraviolet spectrophotometer. Data of the concentration of gene hhl1 is used to calculate the relative expression level of mRNA. PHHL(BBa_K2089004) , PphyB(BBa_K2089003) , Ppif1(BBa_K2089001) are also the promoters we use in our project.
| Growth light | high light | |||
|---|---|---|---|---|
| REL | SE | REL | SE | |
| Phhl1 | 1 | 0.05 | 1.1 | 0.07 |
| Pcop1 | 1 | 0.03 | 10.1 | 1.26 |
| Ppif1 | 1 | 0.06 | 8.9 | 1.01 |
| PphyB | 1 | 0.09 | 15.6 | 1.36 |
This graph shows the relative expression level of mRNA of gene hhl1 expressed by 4 different parts in growth light(100lux) and high light(1200lux).
Pcop1 do not respond to growth light, the expression level of mRNA is similar to that of pHhl1.However, under high light,Pcop1 respond specifically.(The transcription level of Pcop1 under highlight are 10.1 times of the mRNA level under growth light,suggesting that this part can help produce more protein HHL1 to protect PSⅡ under high light
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 96
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 96
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 240
Illegal BamHI site found at 1790 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 96
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 96
- 1000COMPATIBLE WITH RFC[1000]
| None |