Published April 2002
| Published
Journal Article
Open
MALDI-TOF mass spectrometry methods for evaluation of in vitro aminoacyl tRNA production
Abstract
Unnatural amino acid mutagenesis requires the in vitro production of aminoacyl tRNAs. Bacteriophage T4 RNA ligase is used to ligate a-amino-protected dCA amino acids to 74mer tRNA. Previously, there has been no facile method for evaluating the efficiency of this reaction prior to using the tRNA in translation. We report a novel use of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry in monitoring the formation of aminoacyl 76mer tRNA. This method is more efficient and precise than the traditional technique of gel electrophoresis. These MALDI conditions should also prove useful for analyzing aminoacyl tRNAs produced through aminoacyl tRNA synthetases and other methods.
Additional Information
© 2002 RNA Society. Received January 25, 2002; returned for revision February 1, 2002; revised manuscript received February 7, 2002. The authors thank Dr. John F. Leite for his valuable input. This work was supported by the National Institutes of Health (NS-34407 and NS-11756).Attached Files
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Additional details
- PMCID
- PMC1370275
- Eprint ID
- 7718
- Resolver ID
- CaltechAUTHORS:PETrnao02
- NS-34407
- NIH
- NS-11756
- NIH
- Created
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2007-07-19Created from EPrint's datestamp field
- Updated
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2020-03-09Created from EPrint's last_modified field