Purification of eukaryotic translation factors from wheat germ for reconstitution of protein synthesis.
The wheat germ cell-free protein synthesis is a powerful and versatile method for preparation of proteins based on the accumulated DNA sequence information. As the cell extract used for it contains many factors that are unknown or do not directly involve in protein synthesis, details of the translation reaction is yet to be understood. Therefore, we have decided to try reconstitution of protein synthesis, which would be useful for better understanding of the mechanisms supporting eukaryotic protein synthesis and translational regulation and probably applicable to synthetic biology. In the present study, we fractionated an extract from crude wheat germ according to published protocols to obtain the fractions containing the eukaryotic elongation factors (eEFs) 1A, 1B, and 2. The eEF1A and eEF2 fractions supported polyphenylalanine synthesis.
Nagano
Hikaru
H
Department of Material Science and Biotechnology, Graduate School of Science and Engineering, Ehime University, Matsuyama 790-8577, Japan.
Sugihara
Shouhei
S
Takagi
Hisanori
H
Ogasawara
Tomio
T
Endo
Yaeta
Y
Takai
Kazuyuki
K
D002474:Cell-Free System; D020963:GTP Phosphohydrolase-Linked Elongation Factors / Q000302:isolation & purification*; D020648:Peptide Elongation Factor 1 / Q000302:isolation & purification / Q000378:metabolism; D020652:Peptide Elongation Factor 2 / Q000302:isolation & purification / Q000378:metabolism; D010455:Peptides / Q000378:metabolism; D014176:Protein Biosynthesis; D014908:Triticum / Q000737:chemistry / Q000196:embryology
D020648:Peptide Elongation Factor 1; D020652:Peptide Elongation Factor 2; D010455:Peptides; C012001:polyphenylalanine; D020963:GTP Phosphohydrolase-Linked Elongation Factors