title: Protein synthesis directly from PCR: progress and applications of cell-free protein synthesis with linear DNA.
abstract: A rapid, versatile method of protein expression and screening can greatly facilitate the future development of therapeutic biologics, proteomic drug targets and biocatalysts. An attractive candidate is cell-free protein synthesis (CFPS), a cell-lysate-based in vitro expression system, which can utilize linear DNA as expression templates, bypassing time-consuming cloning steps of plasmid-based methods. Traditionally, such linear DNA expression templates (LET) have been vulnerable to degradation by nucleases present in the cell lysate, leading to lower yields. This challenge has been significantly addressed in the recent past, propelling LET-based CFPS as a useful tool for studying, screening and engineering proteins in a high-throughput manner. Currently, LET-based CFPS has promise in fields such as functional proteomics, protein microarrays, and the optimization of complex biological systems.
authors.affiliation: Department of Chemical Engineering, Brigham Young University, Provo, UT, USA.
authors.forename: Song-Min
authors.initials: SM
authors.lastname: Schinn
authors.affiliation: Department of Chemical Engineering, Brigham Young University, Provo, UT, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Broadbent
authors.affiliation: Department of Chemical Engineering, Brigham Young University, Provo, UT, USA.
authors.forename: William T
authors.initials: WT
authors.lastname: Bradley
authors.affiliation: Department of Chemical Engineering, Brigham Young University, Provo, UT, USA. Electronic address: bundy@byu.edu.
authors.forename: Bradley C
authors.initials: BC
authors.lastname: Bundy
mesh_terms: D001709:Biotechnology; D002474:Cell-Free System; D004247:DNA / Q000235:genetics*; D003851:Deoxyribonucleases / Q000037:antagonists & inhibitors; D023281:Genomics; D057166:High-Throughput Screening Assays; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D040081:Protein Array Analysis; D014176:Protein Biosynthesis / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D019151:Peptide Library; D011994:Recombinant Proteins; D004247:DNA; D003851:Deoxyribonucleases