A mini-IRES sequence for stringent selection of high producers
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Abstract
Internal Ribosome Entry Site (IRES) sequences have been widely used to link the expression of two independent proteins on the same mRNA transcript. Genes encoding fluorescent proteins or drug-resistance enzymes are usually placed downstream of IRES, serving as expression indicators or selection markers. In biological applications where the upstream gene-of-interest is to be expressed at extremely high levels, it is often desirable to purposely reduce IRES downstream gene expression to economize the cellular resources and/or to generate more stringent selection pressure. Here we describe a miniature IRES mutant sequence (IRESmut3) with dramatically diminished co-translational efficiency to fulfill these purposes.
Keywords
Co-translation EGFP IRES mutationNotes
Acknowledgements
This work was supported by Juvenile Diabetes Research Foundation (1-2007-551 and 41-2009-760 to WG), and by the National Natural Science Foundation of China (81070624, 81273227), Natural Science Foundation Project of CQ CSTC (2011BB5034) and Foundation for Returned Scholars Sponsored by Department of Human Resources and Social Security of China (CQ-2011-235).
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