Journal
METHODS
Volume 44, Issue 2, Pages 74-80Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2007.08.001
Keywords
Ribosomes; synthetases
Funding
- NCRR NIH HHS [1C06RR018850-01, C06 RR018850] Funding Source: Medline
- NIGMS NIH HHS [R01 GM037552-21, R01-GM37552-21, R01 GM037552] Funding Source: Medline
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The analysis of reactions involving amino acids esterified to tRNAs traditionally uses radiolabeled amino acids. We describe here an alternative assay involving [3'-P-32]-labeled tRNA followed by nuclease digestion and TLC analysis that permits aminoacylation to be monitored in an efficient, quantitative manner while circumventing many of the problems faced when using radiolabeled amino acids. We also describe a similar assay using [3'-P-32]-labeled aa-tRNAs to determine the rate of peptide bond formation on the ribosome. This type of assay can also potentially be adapted to study other reactions involving an amino acid or peptide esterified to tRNA. (C) 2007 Elsevier Inc. All rights reserved.
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