Cell-free chassis/Commercial E. coli T7 S30

Commercial E. coli T7 S30

Cell-Free Systems (back to Intro)	Introduction
Chassis description	The Commercial E. coli T7 S30 extract was purchased from Promega. It was prepared by modifications of the method described by Zubay et al (1980). E. coli strain B deficient in OmpT endoproteinase and lon protease activity was used. The simple constitutive gene expression device BBa_E7104 has been used to characterize this cell-free chassis
Temperature dependence	Graph 1. Graph of GFPmut3b synthesized (pmol) against time (minutes) at 4oC (dark blue), 25oC (pink) and 37oC. The simple constitutive gene expression device BBa_E7104 has been used. Each of the three coloured lines show average measurements based on three replicates of the experiment. The error bars represent the standard deviation of the measurements.
Peak time	Graph 2. Graph of rate of GFPmut3b synthesis (pmol per min) against time (minutes) at 4oC (dark blue), 25oC (pink) and 37oC (yellow). The simple constitutive gene expression device BBa_E7104 has been used. Each of the three coloured lines show average measurements based on three replicates of the experiment. The error bars represent the standard deviation of the measurements.
Expression lifespan	Graph 3. Graph of rate of GFP synthesis (mol per min) against time (minutes) at 4oC (blue), 25oC (red) and 37oC (green). The simple constitutive gene expression device BBa_E7104 has been used. Each of the three coloured lines show average measurements based on three replicates of the experiment. The error bars represent the standard deviation of the measurements.
Expression capacity	Graph 3. Bar chart showing total GFP synthesized in 6 hours (mol) at 4oC (blue), 25oC (red) and 37oC (green). The simple constitutive gene expression device BBa_E7104 has been used. Each of the three coloured bars show average measurements based on three replicates of the experiment. The error bars represent the standard deviation of the measurements. This bar chart chart is derived from Graph 1, with addition data from our GFP calibration curve. Please visit our OpenWetWare for the detailed protocol and raw data of the experiment.

References

Please refer to the [http://www.promega.com/tbs/tb219/tb219.pdf Promega cell extract user guide] for detailed information about the Commercial E. coli T7 S30 extract.