Difference between revisions of "Part:BBa K410000"

Revision as of 18:43, 7 November 2010

Periplasmic Heat Generation

What it is: This part consists of hybB-ompA-AOX1A. HybB is a cold shock promoter. OmpA is a periplasmic targeting sequence, and AOX1A is a gene responsible for thermogenesis in the sacred lotus, a plant. What it does: Induces AOX1A expression in response to a cold shock. Here, we describe a "cold shock" as incubating E. coli between 20-30C for 10-12 hours. The part should produce a thermogenic response raising the temperature of the E. coli's immediate environment (plate or liquid cultures) above ambient temperature.

Usage and Biology BBa_K410000 Usage and Biology Not understood

Characterization

Absolute Temperature Characterization

pSB1A3 containing biobrick part hybB-OmpA-AOX1a was transformed into E. coli BL21. Transformants were grown in 600 mL of LB media(+carbenicillin) at 37 degrees Celcius overnight. The solutions were centrifuged, the soft pellet collected, and resuspended in equivalent amount of fresh LB media(+carbenicillin). The cultures were then placed in a 10 degrees Celcius incubator for 1 hour. 75 mL were pipetted into three different insulated containers and placed in 15, 20 and 25 degrees Celcius incubators for 1 hour acclimation period. Temperatures were recorded at regular intervals using a thermistor. In addition to the pSB1A3 vector containing the biobrick part, a control (pSB1A3 vector with insert hybB-OmpA-mRFP) was also monitored. The graphs shown below illustrate absolute temperature change over time at different incubation temperatures.