Cell-free system for synthesis of proteins derived from cultured mammalian cells
AssigneeRiken
InventorsHiroaki IMATAKA, Satoshi Mikami, Shigeyuki Yokoyama
Life of this patent
Estimating the term…
Abstract
Prepared is an extract composition having an improved protein synthetic activity in a cell-free protein synthesis system using a mammalian cultured cell extract. An eukaryotic translation initiation factor and/or translational regulator are added to a cell-free protein synthesis system comprising an extract prepared from cultured mammalian cells and a template mRNA. These factors are one or more selected from the group consisting of eukaryotic translation initiation factors 4E (eIF4E), 2 (eIF2) and 2B (eIF2B), and eukaryotic translational regulator p97.
Claims21
A process for producing a protein in a cell-free protein synthesis system comprising
- adding a template uncapped mRNA comprising an IRES derived from a virus nucleic acid, eukaryotic translation initiation factors 2 (eIF2) and 2B (eIF2B) to an extract prepared from cultured mammalian cells and comprising 120-240 mM potassium ion, and incubating the mixture thereof.
A process for producing a protein in a cell-free protein synthesis system comprising
- adding a capped mRNA, and eukaryotic translation initiation factors 2 (eIF2) and 2B (eIF2B) to an extract prepared from cultured mammalian cells and comprising 90-150 mM potassium ion, and
- incubating the mixture thereof.
A composition for synthesis of a protein using a cell-free system, comprising an extract of cultured mammalian cells and additional eukaryotic translation initiation factors 2 (eIF2) and 2B (eIF2B).
A kit for cell-free protein synthesis comprising
- i) an extract prepared from cultured mammalian cells;
- ii) isolated eukaryotic transcription initiation factor 2 (eIF2);
- iii) isolated eukaryotic transcription initiation factor 2E (eIF2E);
- iv) isolated eukaryotic protein synthesis regulator p97;
- and optionally
- v) an uncapped mRNA template comprising a IRES sequence;
- wherein each of i) and v) are packaged separately, and wherein ii), iii) and iv) may be packaged separately or as a mixture thereof.
Dependencies are inferred from the claim text.