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Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems
Published on: April 26, 2019
Translation enhancement by optimized downstream box sequences in Escherichia coli and Mycobacterium smegmatis
1Institute of Infectious Diseases and Molecular Medicine, Health Sciences Faculty, University of Cape Town, Observatory, 7925, Cape Town, South Africa. grush@curie.uct.ac.za
Abstract:
The effect on translation of downstream box sequences optimized for binding to Mycobacterium smegmatis and Escherichia coli 16S rRNA in the absence of a Shine-Dalgarno (SD) region was investigated. The relative translational efficiency of each construct in either M. smegmatis or E. coli was determined. Eradication of the SD region in the absence of a downstream box abolished the translation activity. In contrast, optimized downstream box constructs resulted in a 13- and 18-fold increase in protein synthesis, relative to non-optimized DB controls in E. coli and M. smegmatis, respectively.
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