Recombinant tagging system using ribosomal frameshifting to monitor protein expression. Issue 3 (1st November 2012)

• Record Type: Journal Article
• Authors: Han, Se Jong; Cho, Sayeon; Lowehhaupt, Ky; Park, So‐Young; Sim, Sang Jun; Kim, Yang‐Gyun
• Is Part Of: Biotechnology and bioengineering. Volume 110:Issue 3(2013:Mar.)
• Page Start: 898
• Page End: 904
• Publication Date: 2012-11-01
• Subjects: Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.24740
• Languages: English
• ISSNs: 0006-3592

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>Abstract</title> <p>For rapid and accurate quantitation of recombinant proteins during expression and after purification, we introduce a new tagging strategy that expresses both target proteins and limitedly tagged target proteins together in a single cell at a constant ratio by utilizing <italic>cis</italic>‐elements of programmed ‐1 ribosomal frameshifting (‐1RFS) as an embedded device. ‐1RFS is an alternative reading mechanism that effectively controls protein expression by many viruses. When a target gene is fused to the enhanced green fluorescent protein (EGFP) gene with a ‐1RFS element implanted between them, the unfused target and the target‐GFP fusion proteins are expressed at a fixed ratio. The expression ratio between these two protein products is adjustable simply by changing ‐1RFS signals. This limited‐tagging system would be valuable for the real‐time monitoring of protein expression when optimizing expression condition for a new protein, and in monitoring large‐scale bioprocesses without a large metabolic burden on host cells. Furthermore, this strategy allows for the direct measurement of the quantity of a protein on a chip surface and easy application to proteomewide study of gene products. Biotechnol. Bioeng. 2013; 110: 898–904. © 2012 Wiley Periodicals, Inc.</p> </abstract>