Cell-free chassis/Commercial E. coli T7 S30

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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_I719015 has been used to characterize this cell-free chassis

Temperature dependence
T7 Optimum Temp.JPG
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_I719015 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
T7 Peak Time.JPG
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_I719015 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
T7 Expression Lifespan.JPG
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_I719015 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
S30 Expression Capacity.JPG
Graph 4. Bar chart showing total GFPmut3b synthesized in 6 hours (pmol) at 4oC (dark blue), 25oC (pink) and 37oC (yellow). The simple constitutive gene expression device BBa_I719015 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.

References

Please refer to the Promega cell extract user guide for detailed information about the Commercial E. coli T7 S30 extract.