Difference between revisions of "Part:BBa K2089001"
 
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It is a key negative regulator of phytochrome-mediated seed germination and acts by inhibiting chlorophyll biosynthesis, light-mediated suppression of hypocotyl elongation and far-red light-mediated suppression of seed germination, and promoting negative gravitropism in hypocotyls. Light reduces this activity in a phy-dependent manner. The protein preferentially interacts with the Pfr forms of Phytochrome A (PhyA) and Phytochrome B (PhyB), is physically associated with APRR1/TOC1 and is degraded in red (R) and far-red (FR) light through the ubiquitin (ub)-26S proteasome pathway to optimize photomorphogenic development in Arabidopsis. It also negatively regulates GA3 oxidase expression.
 
It is a key negative regulator of phytochrome-mediated seed germination and acts by inhibiting chlorophyll biosynthesis, light-mediated suppression of hypocotyl elongation and far-red light-mediated suppression of seed germination, and promoting negative gravitropism in hypocotyls. Light reduces this activity in a phy-dependent manner. The protein preferentially interacts with the Pfr forms of Phytochrome A (PhyA) and Phytochrome B (PhyB), is physically associated with APRR1/TOC1 and is degraded in red (R) and far-red (FR) light through the ubiquitin (ub)-26S proteasome pathway to optimize photomorphogenic development in Arabidopsis. It also negatively regulates GA3 oxidase expression.
  
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　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.
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===Usage and Biology===
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　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) , Pcop1(BBa_K2089002) , PphyB(BBa_K2089003) are also the promoters we use in our project.
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[[Image:GDSYZX_1.png|thumb|centre|600px]]
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{| class="wikitable" style="width: 50%;text-align: center;margin: auto;"
 +
|-
 +
|
 +
!colspan="2"|Growth light
 +
!colspan="2"|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
 +
|}
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　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).
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　Ppif1 do not respond to growth light, the expression level of mRNA is similar to that of pHhl1.However, under high light,Ppif1 respond specifically.(The transcription level of Ppif1 under highlight are 8.9 times of the mRNA level under growth light,suggesting that this part can help produce more protein HHL1 to protect PSⅡ under high light.
  
 
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Latest revision as of 08:01, 19 October 2016


pif1 promoter from Arabidopsis

It is a key negative regulator of phytochrome-mediated seed germination and acts by inhibiting chlorophyll biosynthesis, light-mediated suppression of hypocotyl elongation and far-red light-mediated suppression of seed germination, and promoting negative gravitropism in hypocotyls. Light reduces this activity in a phy-dependent manner. The protein preferentially interacts with the Pfr forms of Phytochrome A (PhyA) and Phytochrome B (PhyB), is physically associated with APRR1/TOC1 and is degraded in red (R) and far-red (FR) light through the ubiquitin (ub)-26S proteasome pathway to optimize photomorphogenic development in Arabidopsis. It also negatively regulates GA3 oxidase expression.

　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) , Pcop1(BBa_K2089002) , PphyB(BBa_K2089003) are also the promoters we use in our project.

GDSYZX 1.png


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).

　Ppif1 do not respond to growth light, the expression level of mRNA is similar to that of pHhl1.However, under high light,Ppif1 respond specifically.(The transcription level of Ppif1 under highlight are 8.9 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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1467
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]