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| Cell-Free Systems (back to Intro) | Introduction
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| 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
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| 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_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.
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| 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_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.
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| 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_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.
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| Expression capacity | 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.
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References
Please refer to the Promega cell extract user guide for detailed information about the Commercial E. coli T7 S30 extract.
