Team:NCTU Formosa/biobricks

From 2013.igem.org

Biobricks

To strive for the best results, we made a number of experiment. Our team always hang together in spite of many losses.

Parts submitted to the Registry

Loading.....

Brief Information

Please click on the name of the parts for detailed information that is hosted in the Registry website.

Light-regulated system

BBa_K1017726
  • pcya and ho1 are the enzymes needed to convert heme into chromophore phycocyanobiline(PCB), the red light sensor. Since pcya and ho1 are not naturally produced in E.coli, we use Pcons upstream in order to make E.coli continuously expressed them to synthesize PCB.


Nctu pcya ho1.jpg

BBa_K1017301
  • Cph8 is a chimeric light receptor. It is a fusion of the photoreceptor cph1 and the envZ histidine kinase. cph1 is only active when it binds the chromophore phycocyanobiline (PCB).


Nctu cph8.jpg

BBa_K1017781
  • This is the key part to change Ompc promoter intro red promotor. The lacI will inhibit the function of Plac, thus, the original outcomes will be converted into our expected result.


Nctu lacI plac.jpg

BBa_K1017101
  • Ompc promtor, which can sense red light with the presence of cph8. It will be turned on in the dark ,and be turned off in the bright. For the convenient use, we add lacI and lac promotor downstream the biobrick. This part, so called red promotor, can be activated under red light, and inactive in the dark.


Nctu Pred.jpg

Temperature-regulated system

BBa_K1017602
  • By using mGFP as a reporter gene, we can test whether the 37 °C RBS works.


Nctu 37rbs mGFP.jpg

BBa_K1017603
  • In our circuit, this biobrick is the part of Plux's activation when the temperature reaches to 37oC.


Nctu 37rbs luxr.jpg

Small RNA-regulated system

BBa_K1017403
  • The sRNA is the complement of its rRBS. It can regulate the downstream of rRBS in RNA level by binding onto the rRBS when it is transcribed in order to interrupt ribosomes' work. In addition, adding Plux upstream makes the sequence be controlled by luxR/AHL complex.

Nctu plux srna1.jpg

BBa_K1017404
  • The sRNA is the complement of its rRBS. It can regulate the downstream of rRBS in RNA level by binding onto the rRBS when it is transcribed in order to interrupt ribosomes' work.
    Nctu srna2.jpg
BBa_K1017202
  • The sRNA, base pair with target mRNA, including the Shine-Dalgarno sequence. Thus it prevent ribosome from binding to initiate the translation. The rRBS is designed for sRNA perfect binding, and this rRBS is the RBS which can be bound only for our artificial sRNA(BBa_K1017404).

Nctu rbs2.jpg

BBa_K1017811
  • The sequence let us show the efficiency of the sRNA-rRBS binding by expressing the fluorescence with the combination of a sequence providing the transcription of the sRNA.

Nctu pcons rbs2 mrfp.jpg

BBa_K1017401
  • This part, BBa_K1017401, includes our artificial sRNA-1 and rRBS-1. The non-coding small RNA can bind to the Shine-Dalgarno sequence on rRBS-1 by base-pairing. Once the rRBS-1 is blocked, ribosomes cannot bind to it to translate, thus, gene expressions downstream are decreased. Because of specific binding, rRBS-1 can only be bound by sRNA-1. We add Plux upstream, so this part can be regulated by luxR/AHL complex. There is one important thing hasn't be mentioned is that it is a temporary sequence which contains the restriction enzyme cutting sites of SpeI, EcoRI and XbaI in order for us to separate them.

Nctu sRNA rRBS1.jpg

BBa_K1017402
  • This part, BBa_K1017402, is similar to BBa_K1017401 mentioned above, but without Plux. rRBS-2 can only be bound by sRNA-2 due to specificity, then gene expressions downstream are decreased. There is one important thing hasn't be mentioned is that it is a temporary sequence which contains the restriction enzyme cutting sites of SpeI, EcoRI and XbaI in order for us to separate them.

Nctu srna2 rRNS.jpg