Regulatory

Part:BBa_K2089003

Designed by: Rouxian Zeng   Group: iGEM16_GDSYZX-United   (2016-10-11)


phyB promoter from Arabidopsis

Red/far-red photoreceptor involved in the regulation of de-etiolation. Exists in two inter-convertible forms: Pr and Pfr (active). Involved in the light-promotion of seed germination and in the shade avoidance response.

 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) , Ppif1(BBa_K2089001) 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).

 PphyB do not respond to growth light, the expression level of mRNA is similar to that of pHhl1.However, under high light,PphyB respond specifically.(The transcription level of PphyB under highlight are 15.6 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
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 61
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 61
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 61
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 61
  • 1000
    COMPATIBLE WITH RFC[1000]


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