Cell-free protein synthesis on a microchip.
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.
Tabuchi
Marl
M
Department of Medicinal Chemistry, Faculty of Pharmaceutical Sciences, The University of Tokushima, Shomachi, Japan. tabuchi@ph.tokushima-u.ac.jp
Hino
Mami
M
Shinohara
Yasuo
Y
Baba
Yoshinobu
Y
D000818:Animals; D002474:Cell-Free System; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008904:Miniaturization; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis / Q000138:chemical synthesis*; D020543:Proteome / Q000737:chemistry; D014764:Viral Proteins
D011506:Proteins; D020543:Proteome; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases