Stable mammalian cell line expressing a bacteriophage RNA polymerase
AssigneeThe United States of America, as represented by the Secretary, Department of Health and Human Services
InventorsBernard Moss, Orna Elroy-Stein
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Abstract
The present invention relates to a eukaryotic cell line which expresses a foreign RNA polymerase gene. The invention further relates to a method of expressing a foreign protein in a eukaryotic environment utilizing the cell line. The present invention allows for the expression of a foreign protein in a eukaryotic cell without requiring transfecting or infecting the cell with a vector carrying the RNA polymerase gene.
Claims13
A mammalian cell comprising:
- i) a bacteriophage RNA polymerase gene expressing the polymerase wherein the bacteriophage is selected from the group consisting of T7, SP6, GH1, and T3 ;
- ii) a construct comprising the following elements operably linked:
- a) a promoter recognized by the protein product of said bacteriophage RNA polymerase gene;
- b) a DNA sequence that codes for an untranslated region of a piconnavirus RNA that confers cap independent translation;
- and
- c) a gene encoding a protein to be expressed.
A method of expressing a gene in a mammalian cell expressing a bacteriophage RNA polymerase gene, wherein the bacteriophage is selected from the group consisting of T7, SP6, GH1, and T3, which method comprises the steps of:
- introducing into said cell a construct comprising the following elements operably linked:
- a) promoter recognized by the protein product of said RNA polymerase gene;
- b) a DNA sequence that codes for an untranslated region of a cardiovirus RNA that confers cap independent translation;
- and
- c) a gene encoding a protein to be expressed;
- and culturing said cell under conditions such that said gene is expressed.
Dependencies are inferred from the claim text.