Ribosomal synthesis and in situ isolation of peptide molecules in a cell-free translation system.
Although the cell-free translation system is now widely accepted as an efficient platform for production, engineering and screening of recombinant proteins, it has not been successfully used for the synthesis of peptide molecules mainly due to low expression yields and rapid proteolysis of the expressed peptides. In this study, we propose a novel strategy for rapid expression and recovery of peptide molecules which involves the rational design of template DNA and heterogenous cell-free translation reaction in the presence of affinity beads. Various peptide molecules which were not expressed in a detectable level were successfully expressed and recovered in situ in a substantial yield. We expect that the presented approach will be widely used as a versatile platform for the generation of a variety of peptide molecules.
Lee
Kyung-Ho
KH
Interdisciplinary Program for Nano-Technology, Chungnam National University, Daejeon 305-764, Republic of Korea.
Kwon
Yong-Chan
YC
Yoo
Sung Joon
SJ
Kim
Dong-Myung
DM
D001671:Biochemistry / Q000379:methods*; D002474:Cell-Free System; D003062:Codon / Q000235:genetics; D006801:Humans; D055767:Immobilized Proteins / Q000378:metabolism; D008280:Magnetics; D008863:Microspheres; D010455:Peptides / Q000302:isolation & purification*; D014176:Protein Biosynthesis; D021382:Protein Sorting Signals; D055550:Protein Stability; D012270:Ribosomes / Q000378:metabolism*
D003062:Codon; D055767:Immobilized Proteins; D010455:Peptides; D021382:Protein Sorting Signals