Efficient incorporation of a nonnatural amino acid into a protein in an insect cell-free translation system.
Recently, we have succeeded in incorporating various nonnatural amino acids into proteins by using four-base codon-anticodon pairs in Spodoptera frugiperda 21 (Sf21) insect cell-free protein synthesizing system. Here, the reaction was conducted under various conditions in order to optimize the incorporation efficiency. The optimal concentration of aminoacyl-tRNA, reaction temperature, and reaction time were 2 nM, 25 degrees C, and 1.5 hr, respectively.
Tokuda
Yasunori
Y
Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan.
Taki
Masumi
M
Sisido
Masahiko
M
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D000926:Anticodon / Q000737:chemistry; D002474:Cell-Free System; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000378:metabolism; D014176:Protein Biosynthesis; D015202:Protein Engineering / Q000379:methods*; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry / Q000378:metabolism; D018411:Spodoptera / Q000235:genetics; D019809:Streptavidin / Q000096:biosynthesis / Q000737:chemistry; D013696:Temperature
D000596:Amino Acids; D000926:Anticodon; D012346:RNA, Transfer, Amino Acyl; C040239:4-nitrophenylalanine; D010649:Phenylalanine; D019809:Streptavidin