a riboswitch-based inducible gene expression system for mycobacteria一个riboswitch-based分枝杆菌诱导的基因表达系统.pdfVIP

A Riboswitch-Based Inducible Gene Expression System for Mycobacteria 1 2 2 1 5 Jessica C. Seeliger , Shana Topp , Kimberly M. Sogi , Mary L. Previti , Justin P. Gallivan , Carolyn R. Bertozzi2,3,4* 1 Department of Pharmacological Sciences, Stony Brook University, Stony Brook, New York, United States of America, 2 Departments of Chemistry, University of California, Berkeley, California, United States of America, 3 Molecular and Cell Biology, University of California, Berkeley, California, United States of America, 4 Howard Hughes Medical Institute, University of California, Berkeley, California, United States of America, 5 Department of Chemistry and the Center for Fundamental and Applied Molecular Evolution, Emory University, Atlanta, Georgia, United States of America Abstract Research on the human pathogen Mycobacterium tuberculosis (Mtb) would benefit from novel tools for regulated gene expression. Here we describe the characterization and application of a synthetic riboswitch-based system, which comprises a mycobacterial promoter for transcriptional control and a riboswitch for translational control. The system was used to induce and repress heterologous protein overexpression reversibly, to create a conditional gene knockdown, and to control gene expression in a macrophage infection model. Unlike existing systems for controlling gene expression in Mtb, the riboswitch does not require the co-expression of any accessory proteins: all of the regulatory machinery is encoded by a short DNA segment directly upstream of the target gene. The inducible riboswitch platform has the potential to be a powerful general strategy for creating customized gene regulation syst