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KMID : 1094720060110050420
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Biotechnology and Bioprocess Engineering
2006 Volume.11 No. 5 p.420 ~ p.424
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Preparation method for Escherichia coli S30 extracts completely dependent upon tRNA addition to catalyze cell-free protein synthesis
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Ahn Jin-Ho
Hwang Mi-Yeon
Oh In-Seok
Park Kyung-Moon
Hahn Geun-Hee
Choi Cha-Yong
Kim Dong-Myung
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Abstract
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A simple method for depletingE. coli S30 extracts of endogenous tRNA has been developed. An ethanolamine-Sepharose® column equilibrated with water selectively captured the tRNA molecules inE. coli S30 extracts. As a result, S30 extracts filtered through this column became completely dependent upon the addition of exogenous tRNA to mediate cell-free protein synthesis reactions. We anticipate that the procedures developed and described will be particularly useful forin vitro suppression reaction studies designed to introduce unnatural amino acids into protein molecules.
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KEYWORD
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cell-free protein synthesis, unnatural amino acid, in vitro suppression, tRNA, chloramphenicol, acetyltransferase, erythropoietin
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