Analysis of protein functions through a bacterial cell-free protein expression system.
Cell-free protein synthesis is a suitable protein expression method for the high-throughput use because a PCR-amplified linear DNA fragment is utilized as a template for protein synthesis without any cloning procedures. We have developed a two-step PCR method for high-throughput and robust production of linear templates ready for cell-free protein synthesis. A high-throughput protein expression method has been established by combining the batch-mode cell-free protein synthesis with the two-step PCR, which is performed on multiwell plates, and is thus adapted for robotics. In this chapter, our two-step PCR method and the batch-mode cell-free protein synthesis are described.
Kigawa
Takanori
T
RIKEN Systems and Structural Biology Center, Yokohama, Japan.
D000818:Animals; D057205:Automation, Laboratory; D001419:Bacteria / Q000235:genetics / Q000378:metabolism*; D002474:Cell-Free System; D006801:Humans; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D012371:Robotics
D011994:Recombinant Proteins