Composite

Part:BBa_K2089006

Designed by: Rouxian Zeng   Group: iGEM16_GDSYZX-United   (2016-10-12)


pHHL-hhl1

This biobrick consists of a complete functional protein HHL1 coding sequence in Arabidopsis and gene hhl1.Gene hhl1 encodes protein HHL1,which is produced and distributed among chloroplasts(especailly grana core-,grana margin and stroma lamellae enriched thylokoid membranes) to function in protecting and repairing photosystemⅡfrom photodamage. pHhl1 is the promoter of gene hhl1 in wild type Arabidopsis,the promoter may be regulated by the concentration of certain protein differs from before and after photodamage takes place in photosystemⅡ.

 In order to examine the efficiency of this device,we made a part which consists of a complete functional protein HHL1 coding sequence in Arabidopsis and gene hhl1,and this part has been taken into the nucleus of protoplasts of Arabiodopsis using psb1c3 plasmid as its carrier.RT-PCR is done to the transcribed mRNA of gene hhl1 to get cDNA,then we use the ultraviolet spectrophotometer to measure the concentration of cDNA we got.The concentration data is used to calculate the relative expression level of gene hhl1 compared to the gene expression of gene hhl1 regulated by the promoter Ppif1(BBa_K2089001) ,Pcop1(BBa_K2089002) , PphyB(BBa_K2089003).

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

 From the result,this part has no significent change in expression level of mRNA in growth light(100lux) and in high light(1200lux).This means that the promoter pHhl1 is repressed in these light intensities so the expression level is quite low.

Sequence and Features

Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 746
    Illegal PstI site found at 2352
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 187
    Illegal SpeI site found at 746
    Illegal PstI site found at 2352
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 746
    Illegal PstI site found at 2352
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 746
    Illegal PstI site found at 2352
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SpeI site found at 746
    Illegal PstI site found at 2352


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