Recombinant tagging system using ribosomal frameshifting to monitor protein expression

Se Jong Han, Sayeon Cho, Ky Lowehhaupt, So Young Park, Sang Jun Sim, Yang Gyun Kim

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)898-904
Number of pages7
JournalBiotechnology and Bioengineering
Volume110
Issue number3
DOIs
Publication statusPublished - 2013 Mar 1

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Ribosomal Frameshifting
Proteins
Genes
Monitoring
Recombinant proteins
Viruses
Recombinant Proteins
Purification
Reading
Fusion reactions
Equipment and Supplies

Keywords

  • Fluorescence
  • Protein expression
  • Ribosomal frameshifting
  • Tagging system

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Recombinant tagging system using ribosomal frameshifting to monitor protein expression. / Han, Se Jong; Cho, Sayeon; Lowehhaupt, Ky; Park, So Young; Sim, Sang Jun; Kim, Yang Gyun.

In: Biotechnology and Bioengineering, Vol. 110, No. 3, 01.03.2013, p. 898-904.

Research output: Contribution to journalArticle

Han, Se Jong ; Cho, Sayeon ; Lowehhaupt, Ky ; Park, So Young ; Sim, Sang Jun ; Kim, Yang Gyun. / Recombinant tagging system using ribosomal frameshifting to monitor protein expression. In: Biotechnology and Bioengineering. 2013 ; Vol. 110, No. 3. pp. 898-904.
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