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Purification of individual tRNAs using a monoclonal anti-AMP antibody affinity column
Authors:R X Zhu  W M Ching  H K Chung  S G Rhee  T C Stadtman
Affiliation:1. Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD 20892, USA;1. Institute of Applied Computer Science, Lodz University of Technology, Stefanowskiego 18/22, 90-924 Lodz, Poland;2. Department of Plant Physiology and Biochemistry, Faculty of Biology and Environmental Protection, University of Lodz, Banacha 12/16, 90-237 Lodz, Poland;1. Department of Medical Laboratory, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, Guangdong 510515, China;2. Center for Translational Medicine, Precision Medicine Institute, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510080, China;3. State Key Laboratory of Oncology in South China, Sun Yat-sen University Cancer Center, Guangzhou, Guangdong 510080, China;1. Department of Plant Biology and Genome Center, University of California, Davis, Davis, CA 95616, USA;2. Plant Ecophysiology, Institute of Environmental Biology, Utrecht University, 3584 Utrecht, the Netherlands;3. Plant Biology Graduate Group, University of California, Davis, Davis, CA 95616, USA;4. Integrative Genetics and Genomics Graduate Group, University of California, Davis, Davis, CA 95616, USA;5. Center for Plant Cell Biology, Department of Botany and Plant Sciences, University of California, Riverside, Riverside, CA 92521, USA;6. IBBM, FCE-UNLP CONICET, La Plata 1900, Argentina;7. Department of Cell and Systems Biology/Centre for the Analysis of Genome Evolution and Function, University of Toronto, 25 Willcocks St., Toronto, ON M5S 3B2, Canada;8. Department of Plant Biology, University of California, Davis, Davis, CA 95616, USA;9. Fort Valley State University, Fort Valley, GA 31030, USA;10. Department of Biomedical Engineering and Genome Center, University of California, Davis, Davis, CA 95616, USA;11. Department of Biology, Emory University, Atlanta, GA 30322, USA;12. Department of Biology, University of Padova, Padova, Italy;13. Department of Plant Sciences, University of California, Davis, Davis, CA 95616, USA;14. Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, 1432 Ås, Norway;1. Laboratory of Molecular Modelling and Drug Discovery, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genoa, Italy;2. European Molecular Biology Laboratory Grenoble, 71 Avenue des Martyrs, Grenoble 38042, France
Abstract:A murine monoclonal anti-AMP antibody affinity matrix was used for isolation of individual species of amino acid transfer nucleic acids (tRNAs). The antibodies had been prepared using 5'-AMP covalently attached to bovine serum albumin as antigen and exhibited high affinity for 5'-AMP but greatly reduced affinity for 3'-AMP. Native uncharged tRNAs that terminate in a 5'-AMP group on the amino acid acceptor arm of the molecule bind tightly to the anti-AMP affinity matrix, whereas aminoacylated tRNAs are not retained. This allows separation of a particular tRNA species as its aminoacyl derivative from a complex mixture of uncharged tRNAs under very mild conditions.
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