Cell-free protein synthesis on a microchip

• Published in: Proteomics (Weinheim) 2002-04, Vol.2 (4), p.430-435
• Main Authors: Tabuchi, Mari, Hino, Mami, Shinohara, Yasuo, Baba, Yoshinobu
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Cell-free protein synthesis; Cell-Free System; Cloning vector; Diverse techniques; DNA-Directed RNA Polymerases - chemistry; DNA-Directed RNA Polymerases - genetics; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Humans; Microchip; microchips; Miniaturization; Molecular and cellular biology; Phage T7; Protein Array Analysis - instrumentation; Protein Array Analysis - methods; Protein Biosynthesis; Proteins; Proteins - analysis; Proteins - chemical synthesis; Proteome; Proteome - chemistry; proteomics; Viral Proteins
• ISSN: 1615-9853; 1615-9861
• DOI: 10.1002/1615-9861(200204)2:4<430::AID-PROT430>3.0.CO;2-5

We evaluated the expression of various proteins by using a cell‐free protein synthesis system with a rapid translation system, a microtube, and a microchip technique. Protein expression was successfully achieved with a microfabricated reaction chamber on a plastic chip. Proteins were expressed effectively by use of expression vectors for T7 RNA polymerase (pET) instead of a plasmid for in vitro expression vector, which is recommended by the manufacture of the rapid translation system. Expression of the proteins depended on the type of proteins chosen. Two mammalian proteins were synthesized simultaneously with two expression vectors of pET species. Effective application of the cell‐free protein synthesis system will enable miniaturization of protein synthesis and mediation between the transcriptome and proteome.