Fluorescent labeling of cell-free synthesized proteins with fluorophore-conjugated methionylated tRNA derived from in vitro transcribed tRNA
- Authors
- Jun, Soo Youn; Kang, Sang Hyeon; Lee, Kwang-Ho
- Issue Date
- Jun-2008
- Publisher
- ELSEVIER SCIENCE BV
- Keywords
- cell-free synthesized proteins; fluorescence; labeling; in vitro transcription
- Citation
- JOURNAL OF MICROBIOLOGICAL METHODS, v.73, no.3, pp 247 - 251
- Pages
- 5
- Journal Title
- JOURNAL OF MICROBIOLOGICAL METHODS
- Volume
- 73
- Number
- 3
- Start Page
- 247
- End Page
- 251
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/23727
- DOI
- 10.1016/j.mimet.2008.02.016
- ISSN
- 0167-7012
1872-8359
- Abstract
- A simple and practical method for preparing fluorophore-conjugated methionylated tRNA necessary for tRNA-mediated fluorescent labeling of cell-free synthesized proteins was developed. Without complicated chromatographic purification and subsequent concentration, fluorophore-conjugated methionylated tRNA with higher purity and fluorescence concentration could be synthesized from in vitro transcribed tRNA instead of from a total IRNA mixture, which has been routinely used as a tRNA source. Although fluorophore-conjugated methionylated tRNA derived from in vitro transcribed tRNA was purified by simple phenol extraction following alcohol precipitation, it worked well in tRNA-mediated fluorescent labeling, yielding an improved signal-to-noise ratio and higher fluorescence intensity compared to the conventional total tRNA-based method. Based on its simplicity in the preparation of labeling agent with higher purity and fluorescence concentration, the developed method will accelerate the prevalence of fluorescence-based detection of cell-free synthesized proteins. (C) 2008 Elsevier B.V. All rights reserved.
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Collections - College of Natural Sciences > Department of Life Science > 1. Journal Articles
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