RNA Scaffolds pp 105-112 | Cite as

RNA Scaffold: Designed to Co-localize Enzymes

Part of the Methods in Molecular Biology book series (MIMB, volume 1316)

Abstract

Self-assembling RNA scaffold is designed to co-localize enzymes in engineered biological pathways through interactions between scaffold’s protein docking domains and their affinity protein–enzyme fusions, in vivo. Here we introduced a noncoding RNA structure theophylline aptamer that respond to theophylline ligand in order to modulate RNA scaffold and regulate multistep enzymatic activities. We described the specifically designed RNA scaffold increased the fluorescent intensity in a splitting GFP assay by 2.25-fold, and also observed a 1.43-fold increase in multi-enzymatic efficiency in IAA synthesis pathway.

Key words

RNA scaffold Aptamer Riboswitch Splitting GFP Co-expression Overlap PCR 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.College of Life SciencesZhejiang UniversityHangzhouChina

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