An In Vitro Assay to Screen for Translation Inhibitors

• Published in: Journal of microbiology and biotechnology Vol. 16; no. 10; pp. 1646 - 1649
• Main Authors: Song, C.H. (Sunchon National University, Sunchon, Republic of Korea), Paik, H.R. (Sunchon National University, Sunchon, Republic of Korea), Seong, C.N. (Sunchon National University, Sunchon, Republic of Korea), Choi, S.K. (Sunchon National University, Sunchon, Republic of Korea), E-mail: sangkic@sunchon.ac.kr
• Format: Journal Article
• Language: English
• Published: Seoul Korean Society for Applied Microbiology 01.10.2006; 한국미생물·생명공학회
• Subjects: Biological and medical sciences; Biotechnology; Fundamental and applied biological sciences. Psychology; In vitro translation; inhibitor; luciferase; PROTEIN SYNTHESIS; SINTESIS DE PROTEINAS; STREPTOMYCES; SYNTHESE PROTEIQUE; 생물학; Streptomycetaceae; In vitro translation; Enzyme; Luciferase; Streptomyces; In vitro; Genetic translation; Actinomycetales; Protein synthesis; Streptomyces spp; Bacteria; Actinomycetes; Inhibitor; Oxidoreductases
• ISSN: 1017-7825

Protein synthesis is the ultimate outcome of gene expression which, in turn, is regulated by several translation factors. We attempted to identify substances that can inhibit the translation process in vitro when the outcome protein is luciferase. To this end, we developed a sensitive cell-free protein synthesis assay using luciferase as the reporter. The synthesis of luciferase increased proportionately as mRNA was added to a 15-μl reaction medium in concentrations raging from 5 ng to 500 ng. The maximum amount of luciferase was synthesized when the media were incubated at 25℃ for 40 min.