caa a22 4500 397555253 CHVBK 20180308164759.0 cr unu---uuuuu 161202e199602 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021231 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021231 A Method for Screening Antisense Oligodeoxyribonucleotides Effective for mRNA Translation-Arrest [Elektronische Daten] [Trong-Zong Chen, Shwu-Bin Lin, Jaw-Ching Wu, Kong-Bung Choo, Lo-Chun AuI] A transcription and translation coupled reticulocyte lysate system was established for rapid screening of antisense oligodeoxyribonucleotides (ODNs) to determine which are most effective for mRNA translation-arrest. A plasmid containing the target cDNA under the control of the T7 (or SP6) promoter was added to the lysate system in the presence of the T7 (or SP6) RNA polymerase, RNase H, and the antisense ODN under test. Transcription and translation were accomplished in a one-tube reaction. Translation-arrest caused by antisense ODN was evaluated in terms of the amounts of de novo-synthesized, [35S]-methionine or [35S]cysteine labeled target protein measured by gel electrophoresis and autoradiography. The properties of this system and optimal reaction conditions for use in antisense ODN screening were determined. Our method is simpler and more rapid than other in vitro screening methods. © by the Journal of Biochemistry Regular Papers nationallicence antisense oligonucleotide nationallicence de novo synthesis nationallicence in vitro screening nationallicence transcription and translation coupled system nationallicence translation-arrest nationallicence Chen Trong-Zong Institute of Clinical Medicine, School of Medicine Taiwan aut Lin Shwu-Bin Graduate Institute of Medical Technology, College of Medicine, National Taiwan University Veterans General Hospital- Taipei, Taipei, Taiwan aut Wu Jaw-Ching Institute of Clinical Medicine, School of Medicine Taiwan aut Choo Kong-Bung Graduate Institute of Medical Biotechnology, National Yang-Ming University Taiwan aut AuI Lo-Chun Graduate Institute of Medical Biotechnology, National Yang-Ming University Taiwan aut The Journal of Biochemistry Oxford University Press 119/2(1996-02), 252-255 0021-924X 119:2<252 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021231 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021231 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Chen Trong-Zong Institute of Clinical Medicine, School of Medicine Taiwan aut NATIONALLICENCE 700 1- Lin Shwu-Bin Graduate Institute of Medical Technology, College of Medicine, National Taiwan University Veterans General Hospital- Taipei, Taipei, Taiwan aut NATIONALLICENCE 700 1- Wu Jaw-Ching Institute of Clinical Medicine, School of Medicine Taiwan aut NATIONALLICENCE 700 1- Choo Kong-Bung Graduate Institute of Medical Biotechnology, National Yang-Ming University Taiwan aut NATIONALLICENCE 700 1- AuI Lo-Chun Graduate Institute of Medical Biotechnology, National Yang-Ming University Taiwan aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/2(1996-02), 252-255 0021-924X 119:2<252 1996 119 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford