title: Recombinant tagging system using ribosomal frameshifting to monitor protein expression.
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.
authors.affiliation: Division of Polar Life Sciences, Korea Polar Research Institute, Incheon, Korea.
authors.forename: Se Jong
authors.initials: SJ
authors.lastname: Han
authors.forename: Sayeon
authors.initials: S
authors.lastname: Cho
authors.forename: Ky
authors.initials: K
authors.lastname: Lowehhaupt
authors.forename: So-Young
authors.initials: SY
authors.lastname: Park
authors.forename: Sang Jun
authors.initials: SJ
authors.lastname: Sim
authors.forename: Yang-Gyun
authors.initials: YG
authors.lastname: Kim
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D005453:Fluorescence; D018965:Frameshifting, Ribosomal; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins