Difference between revisions of "GDSYZX-United/PARTS/parts"

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Revision as of 13:56, 8 October 2016

Parts

Part1:PHhl1-hhl1-flu

This biobrick consists of a complete functional protein HHL1 coding sequence in Arabidopsis ,gene hhl1,fused with a flu sequence as a reporter.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,this biobrick 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 part2,3,4.

From the result,part1 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.

Part2:pPhyB-hhl1-flu

This biobrick consists of a photosensitive promoter pPhyB, and protein coding sequence hhl1 fused with gene flu as a reporter. pPhyB is the promoter of gene phyB, which encodes phytochrome B. PhytochromeB is a photoreceptor which is sensitive to red/far-red light(wavelength:660-730nm).This biobrick is constructed to produce more protein HHL1 in chloroplasts to help protect photosystemⅡ before photodamage takes place.

Part3:pPif1-hhl1-flu

This biobrick consists of a photosensitive promoter pPif1,and protein coding sequence hhl1 fused with gene flu as a reporter.This biobrick is constructed to produce more protein HHL1 in chloroplasts to help protect photosystemⅡ before photodamage takes place.

Part4:pCop1-hhl1-flu

This biobrick consists of a photosensitive promoter pCop1,and protein coding sequence hhl1 fused with gene flu as a reporter.This biobrick is constructed to produce more protein HHL1 in chloroplasts to help protect photosystemⅡ before photodamage takes place.