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Experiment | Experiment | ||
<h1>Overview of experiment</h1> | <h1>Overview of experiment</h1> | ||
− | In our project, we first designed toehold switches the detect H7N9 and H5N1 virus in silico based on our modelling. We then constructed the plasmids that express switches and triggers by DNA synthesis and standard cloning method. Validation of the toehold switches was performed by co-expressing the switch and trigger plasmids in E. coli and in cell free system. Meanwhile, we also constructed our toehold switches and trigger cloning tool to allow convenient construction of switch and trigger expressing plasmid. Using the cloning tool, we tried to improve one existing toehold switch in the Registry. We also characterized two reporter protein (mRFP and amaJlime) in the registry to facilitate our project. | + | <font size="4">In our project, we first designed toehold switches the detect H7N9 and H5N1 virus in silico based on our modelling. We then constructed the plasmids that express switches and triggers by DNA synthesis and standard cloning method. Validation of the toehold switches was performed by co-expressing the switch and trigger plasmids in E. coli and in cell free system. Meanwhile, we also constructed our toehold switches and trigger cloning tool to allow convenient construction of switch and trigger expressing plasmid. Using the cloning tool, we tried to improve one existing toehold switch in the Registry. We also characterized two reporter protein (mRFP and amaJlime) in the registry to facilitate our project. |
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− | SDS–PAGE analysis of purification of amajLime (left) and mRFP (right) proteins by ion-exchange chromatography and followed by hydrophobic-interaction chromatography HIC. F.1 to F.3 represents chronological order of elution in HIC. Sample were mixed with 10 µl SDS 2X gel-loading buffer and 10 µl were loaded on the SDS-Gel. The purest fraction, F.2 in both cases, were selected to proceed for pH stability test. | + | SDS–PAGE analysis of purification of amajLime (left) and mRFP (right) proteins by ion-exchange chromatography and followed by hydrophobic-interaction chromatography HIC. F.1 to F.3 represents chronological order of elution in HIC. Sample were mixed with 10 µl SDS 2X gel-loading buffer and 10 µl were loaded on the SDS-Gel. The purest fraction, F.2 in both cases, were selected to proceed for pH stability test.</font size> |
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Revision as of 08:17, 14 October 2017