Part:BBa_K115002:Experience
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Applications of BBa_K115002
The parts were tested in luciferase constructs, by measuring luciferase activity after growing at certain temperatures. The fold increase of luciferase activity between cultures of cells grown at 20ºC and 37ºC is displayed in figure 1.
In this figure, 12 (the leftmost) is construct K115012, the reference strain, which has no thermosensitive RNA, but just B0032 as ribosome binding site. This is to compare for normal temperature induced behaviour in cells. The other constructs from 29-36 are K115029 to K115036, please look at the experience tab of these constructs to see which thermometer RNA they contain.
For more documentation, visit our wiki [http://2008.igem.org/Team:TUDelft (link)].
2010IGEM-TEAM NCTU_formosa's application
We using this parts as a regulator in our project's low-temperature release system. [[1]] [[2]] [[3]]
We constructed series circuits' test as following article.
First, we made A+B circuit in the PSB3K3 plasmid.
in this way we can use it to test by Using Flow Cytometry to obtain the data of A+B
(for40°C, 37°C, 30°C, and 25°C)
and the following figures is our testing results. To test the efficiency of the temperature induced RBS and the interaction between tetR gene and ptet promoter, we replaced the crystal protein gene with GFP (green fluorescence protein) gene. In this scenario, GFP will simulate the crystal protein’s production, as it is easily detected by flow cytometry.
To conclude, we have verified that our strategy works. It demonstrates the regulation of GFP in a temperature dependent fashion, as it is evident the mean GFP values are significantly lower at T>37° C.
We expect our circuit design to allow steady bacteria growth and inhibit crystal protein at T>37° C, and high protein production and low bacteria growth rate at T<37° C
User Reviews
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