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Recombinant tagging system using ribosomal frameshifting to monitor protein expression
Author(s) -
Han Se Jong,
Cho Sayeon,
Lowehhaupt Ky,
Park SoYoung,
Sim Sang Jun,
Kim YangGyun
Publication year - 2013
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.24740
Subject(s) - green fluorescent protein , fusion protein , recombinant dna , gene expression , target protein , ribosomal protein , gene , protein expression , protein biosynthesis , biology , luciferase , microbiology and biotechnology , computational biology , chemistry , ribosome , transfection , biochemistry , rna
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 cis ‐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.