Patent ID: 6635478
Filing Date: 2003-10-21
Classification: C07K,C12N,G01N

Abstract:
A method of producing a mammalian cell line for stable expression of a gene product, comprising the steps of:cotransfecting a mammalian cell with a first construct which cause said cell to express herpes simplex virus transactivating protein VP16 and a second construct comprising a selectable resistance gene to a first antibiotic; selecting said cell which is resistant to the first antibiotic; screening said cell which is resistant to the first antibiotic for VP16 expression; cotransfecting said cell which expresses herpes simplex virus transactivating protein VP16 with third and fourth constructs, the third construct comprising a herpes simplex virus gene promoter that responds to VP16 operably linked to a gene of interest, and the fourth construct comprising a selectable resistance gene to a second antibiotic; selecting said cell which is resistant to the second antibiotic; screening said cell which is resistant to the second antibiotic for expression of the gene product of said gene of interest; and culturing said cell into a stable cell line.