Difference between revisions of "Part:BBa M50477:Design"

Revision as of 01:33, 12 December 2018

Heat-shock-indued chromoprotein expression plasmid (pHeat)

Design Notes

Our pHeat construct begins with heat shock promoter, hsp70 , that will induce the expression of red chromoprotein, meffRed, at higher temperatures. To better quantify the amount of meffRed produced during experimentation, we included a 6x-His tag on the meffRed chromoprotein, in order to allow for Western blot analyses, if necessary. hsp70 is a constitutive heat-shock promoter listed as part BBa_K1895002. 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 (part BBa_K1033922) is a bright red chromoprotein that comes from the coral Montipora efflorescens. It was chosen because of its relatively short length and the fact that it already contains a ribosome binding site.

Basic Parts

Hsp70 promoter: BBa_K1895002

meffRed: BBa_K1033922

6x-His tag: BBa_K823035

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