Team:SYSU-China/Project

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Project Description

Content

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Current methods of high-affinity protein selection are time-consuming and laborious, which hinders the application process. This year, by constructing an integrated protein evolution machine, we made it automatic and efficient. A broad diversification was accomplished by a DNA mutagenesis module in the host bacteria, which enabled prototype protein sequence, carried by the budding-deficient M13 bacteriophage, to generate a library of candidate proteins. Subsequently, bacterial-two-hybrid system was used to give selection pressure on these candidates. Only the favorable candidate proteins can activate compensating gene transcription and rescue phage budding, thus enriching favorable protein sequence in the phage population. Moreover, an RNA thermometer, on the compensating gene mRNA, was applied in the phage amplification process to make it more controllable. Finally, by integrating all the diversification, selection and amplification process, our integrated directed evolution machine can make it an automatic and controllable process for high-affinity protein selection.


References

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