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Preparation method for Escherichia coliS30 extracts completely dependent upon tRNA addition to catalyze cell-free protein synthesis
- Authors
- Ahn, Jin-Ho; Hwang, Mi-Yeon; Oh, In-Seok; Park, Kyung-Moon; Hahn, Geun-Hee; Cho, Cha-Yong; Kim, Dong-Myung
- Issue Date
- Sep-2006
- Publisher
- KOREAN SOC BIOTECHNOLOGY & BIOENGINEERING
- Keywords
- cell-free protein synthesis; unnatural amino acid; in vitro suppression; tRNA; chloramphenicol; acetyltransferase; erythropoietin
- Citation
- BIOTECHNOLOGY AND BIOPROCESS ENGINEERING, v.11, no.5, pp.420 - 424
- Journal Title
- BIOTECHNOLOGY AND BIOPROCESS ENGINEERING
- Volume
- 11
- Number
- 5
- Start Page
- 420
- End Page
- 424
- ISSN
- 1226-8372
- Abstract
- A simple method for depleting E. coli S30 extracts of endogenous tRNA has been developed. An ethanolamine-Sepharose (R) column equilibrated with water selectively captured the tRNA molecules in E. 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 for in vitro suppression reaction studies designed to introduce unnatural amino acids into protein molecules.
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Related Researcher
- Park, Kyung moon
- Science & Technology (Biological and Chemical Engineering)