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30 bacterial extract. Cell-free expression has several advantages compared to conventional recombinant production. The technique is much faster than recombinant production, as the transform Link 1
Cell-free expression is an emerging technique for the small-scale production of recombinant proteins, utilizing an in vitro transcription and translation system such as a Sation or transfection steps can be skipped. Purification is also much easier, as no cells have to be disrupted. Proteins that are normally toxic to cells can easily be expressed. Furthermore, important reaction components can simply be added without the need to traverse the cellular membrane. Moreover, interfering endogenous materials can be removed from the reaction mixture before cell-free expression. One particularly interesting application of cell-free expression is called in vitro compartmentalization1, a technique that can be used to evolve proteins. Our interest lies in the directed evolution of specific viral coat proteins (HaPyV VP12), which can spontaneously form virus-like particles (VLPs) upon overexpression in bacteria (see attached PDF). These VLPs are known to encapsidate dsDNA in a sequence-independent fashion. This allows us to produce transgene-containing virions suitable for gene therapy.
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