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MALDI-TOF mass spectrometry methods for evaluation of in vitro aminoacyl tRNA production

Petersson, E. J. and Shahgholi, M. and Lester, H. A. and Dougherty, D. A. (2002) MALDI-TOF mass spectrometry methods for evaluation of in vitro aminoacyl tRNA production. RNA, 8 (4). pp. 542-547. ISSN 1355-8382. PMCID PMC1370275. https://resolver.caltech.edu/CaltechAUTHORS:PETrnao02

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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.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1370275/PubMed CentralArticle
http://www.rnajournal.org/cgi/content/abstract/8/4/542OtherUNSPECIFIED
http://www.rnajournal.org/cgi/content/abstract/8/4/542OtherUNSPECIFIED
ORCID:
AuthorORCID
Petersson, E. J.0000-0003-3854-9210
Lester, H. A.0000-0002-5470-5255
Dougherty, D. A.0000-0003-1464-2461
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).
Funders:
Funding AgencyGrant Number
NIHNS-34407
NIHNS-11756
Subject Keywords:aminoacylation; gel electrophoresis; matrix-assisted laser desorption/ionization; nonsense suppression; T4 RNA ligase; T7 RNA polymerase; THG73 tRNA; unnatural amino acid
Issue or Number:4
PubMed Central ID:PMC1370275
Record Number:CaltechAUTHORS:PETrnao02
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:PETrnao02
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:7718
Collection:CaltechAUTHORS
Deposited By: Lindsay Cleary
Deposited On:19 Jul 2007
Last Modified:09 Mar 2020 13:19

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