Streamlining the production of proteins for structural biology

摘要

There has been a transformation in the power and throughput of analytic methods delivered by large-scale facilities, such as synchrotrons. This has placed an increasing demand on the supply of high-quality samples (purified proteins and protein crystals) for structural studies. To meet this requirement, protein production has been streamlined to improve efficiency and throughput. Nonetheless, delivering recombinant proteins in sufficient quantity and quality remains a significant bottleneck in the gene to structure pipeline for a variety of reasons including low levels of expression, poor solubility, and toxicity when over-expressed. Many of these issues had been encountered by participants on the START workshop on "Biophysics Structural Biology at Synchrotrons," during the course of their projects. Therefore, our aim over the first day of the workshop was to share our experience of setting up and running protein production workflows that address the challenge of producing a wide variety of proteins for structural analysis. Lessons learnt were explained and some examples of how to clone, express, and purify different proteins were discussed with the participants in some "virtual" practical work and case studies.