Coupled transcription and translation in eukaryotic cell-free extract
AssigneePromega Corporation
InventorsDavid V. Thompson, Thomas R. Van Oosbree, Gregory S. Beckler, John F. Van Herwynen
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Abstract
A method for coupling transcription and translation from DNA using a solution comprising a eukaryotic cell-free extract comprising adding a sufficient amount of a magnesium compound to the extract to raise the magnesium concentration to a level where RNA is transcribed from DNA and RNA translates into protein.
Claims17
A method for coupling transcription and translation in a eukaryotic cell-free extract to produce protein in a static reaction comprising the steps of:
- adding a DNA template to the extract;
- adding ribonucleotides triphosphates to the extract;
- adding a RNA polymerase to the extract;
- and
- adding a sufficient amount of a magnesium salt to the extract to raise the magnesium concentration to a level where RNA is transcribed from the DNA template and RNA translates into protein.
A process for producing protein from a DNA template through coupled transcription and translation in a batch reaction, said process comprising the steps of
- preparing a eukaryotic cell-free extract,
- adding to the extract the DNA template,
- adding to the extract ribonucleotide triphosphates, amino acids and a polymerase, and
- adding a sufficient amount of a magnesium salt to the extract to raise the final magnesium concentration to a level where RNA is transcribed from the DNA template and RNA translates into protein.
A kit for producing protein from a DNA template through coupled transcription and translation, said kit comprising the following components adapted to be used in a batch reaction:
- eukaryotic cell-free extract,
- ribonucleotide triphosphates,
- RNA polymerase, and
- magnesium at a concentration whereby RNA is transcribed from the DNA template and RNA translates into protein.
A eukaryotic cell-free extract for producing protein from a DNA template through coupled transcription and translation in a batch reaction, said extract including ribonucleotide triphosphates, RNA polymerase, and a sufficient amount of a magnesium to raise the final magnesium concentration in the extract to a level where RNA is transcribed from the DNA template and RNA translates into protein.
Dependencies are inferred from the claim text.