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Updated: May 11, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
Published on: March 25, 2020
A mini-IRES sequence for stringent selection of high producers
Jun Yan1, Hailin Yang, Guohua Yue
1Institute of Surgery Research, State Key Laboratory of Trauma, Burns and Combined Injury, Research Institute for Traffic Medicine, Daping Hospital, Third Military Medical University, Chongqing 400042, China.
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.
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