Fluorescence-based Monitoring of the Ribosomal Activity to Optimize Yield from Recombinant Proteins
In vitro transcription/translation systems (ivtt) are used in biotechnology mainly to produce recombinant proteins whose production in vivo would be toxic. The experimental analysis normally involves autoradiography, i.e. it requires the labelling of translation products with radioactive isotopes. Handling these isotopes is expensive, a potential health hazard and limits throughput.
The present invention consists of stable bacterial strains with ribosomal subunits incorporating fluorescent markers, which have growth characteristics similar to wild type and which have an intact translation apparatus. This opens the opportunity for measuring the translation activity in real time using fluorescence. It is possible to carry out experiments in a shorter time and at less cost in multi-well plates, varying several reaction parameters in parallel to optimize yield.
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