Part:BBa_M50477:Design
Heat-shock-indued chromoprotein expression plasmid (pHeat)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 547
Design Notes
Our pHeat construct begins with heat shock promoter, hsp70 (BBa_K1895002) that will induce the expression of red chromoprotein, meffRed (BBa_K1033922), at higher temperatures. We chose this specific heat-shock promoter because it has been the most studied and has been shown to be more effective at sensing hot temperatures. meffRed is a bright red chromoprotein that comes from the coral Montipora efflorescens; this part already contain an RBS. It was chosen because of its relatively short length and the fact that it already contains a ribosome binding site.To better quantify the amount of meffRed produced during experimentation, we included a 6x-His tag (BBa_K112703) on the meffRed chromoprotein, in order to allow for Western blot analyses, if necessary.
Basic Parts
Hsp70 promoter: BBa_K1895002
meffRed: BBa_K1033922
6x-His tag: BBa_K112703
References
Agashe, V. R., & Hartl, F. (2000). Roles of molecular chaperones in cytoplasmic protein folding. Seminars in Cell & Developmental Biology, 11(1), 15-25.
Arsène F, Tomoyasu T, Bukau B. The heat shock response of Escherichia coli. Int J Food Microbiol. 2000 Apr 10;55(1-3):3-9.
Rodriguez JL, et al. 2014. Selection of Escherichia coli heat shock promoters toward their application as stress probes. J Biotechnol. 188:61-71.
Stanford Location
Plasmid name: pHeat
Organism: E. coli
Device type: Sensor
Glycerol stock barcode: 0133012020
Box label: BioE44 F18