Novel plamid DNA comprising reoprter gene DNA and use of the same

Abstract

A plasmid DNA comprising a novel reporter gene DNA; a transformant transformed with the above plasmid DNA and a DNA encoding a known effector protein; a therapeutic agent for a cancer or an autoimmune disease, comprising both of the above DNAs as active ingredients; and a method for detecting a ligand of an intracellular receptor, comprising using the above transformant.

1 - 4 . (canceled)  
     
     
         5 . A transformant transformed with: 
 a plasmid DNA encoding an amino acid sequence comprising a transmembrane region and a functional region of a Fas antigen, in a region downstream of a responsive element of Gal4 protein and,    a DNA encoding an effector protein.    
     
     
         6 . The transformant according to  claim 5 , wherein the effector protein is a fusion protein in which an amino acid sequence comprising a ligand binding region of a nuclear receptor is linked to the C-terminal of an amino acid sequence comprising a DNA binding region of Gal4 protein.  
     
     
         7 . The transformant according to  claim 6 , wherein the nuclear receptor is a steroid receptor.  
     
     
         8 . The transformant according to  claim 6 , wherein the nuclear receptor is a PPAR α receptor.  
     
     
         9 . The transformant according to  claim 8 , wherein the amino acid sequence comprising the ligand binding region of the nuclear receptor is an amino acid sequence of the 167th to 468th positions of human PPAR α receptor or an amino acid sequence of the 167th to 468th positions of mouse PPAR α receptor.  
     
     
         10 . The transformant according to  claim 6 , wherein the nuclear receptor is a PPAR δ receptor.  
     
     
         11 . The transformant according to  claim 10 , wherein the amino acid sequence comprising the ligand binding region of the nuclear receptor is an amino acid sequence of the 139th to 441st positions of human PPAR δ receptor or an amino acid sequence of the 138th to 440th positions of mouse PPAR δ receptor.  
     
     
         12 . The transformant according to  claim 6 , wherein the nuclear receptor is a PPAR γ receptor.  
     
     
         13 . The transformant according to  claim 12 , wherein the amino acid sequence comprising the ligand binding region of the nuclear receptor is an amino acid sequence of the 176th to 478th positions of human PPAR γ1 receptor, an amino acid sequence of the 174th to 475th positions of mouse PPAR γ1 receptor, an amino acid sequence of the 204th to 506th positions of human PPAR γ2 receptor, or an amino acid sequence of the 204th to 505th positions of mouse PPAR γ2 receptor.  
     
     
         14 . The transformant according to  claim 6 , wherein the nuclear receptor is a retinoid X receptor.  
     
     
         15 . The transformant according to  claim 14 , wherein the amino acid sequence comprising the ligand binding region of the nuclear receptor is an amino acid sequence of the 225th to 462nd positions of human retinoid X receptor α, an amino acid sequence of the 297th to 526th positions of human retinoid X receptor β, an amino acid sequence of the 230 to 467th positions of mouse retinoid X receptor α, an amino acid sequence of the 171st to 410 positions of mouse retinoid X receptor β, or an amino acid sequence of the 229th to 463rd positions of mouse retinoid X receptor γ.  
     
     
         16 . The transformant according to  claim 6 , wherein the nuclear receptor is a retinoic acid receptor.  
     
     
         17 . The transformant according to  claim 16 , wherein the amino acid sequence comprising the ligand binding region of the nuclear receptor is an amino acid sequence of the 198th to 462nd positions of human retinoic acid receptor α, an amino acid sequence of the 191 st to 448th positions of human retinoic acid receptor β, an amino acid sequence of the 200th to 454th positions of human retinoic acid receptor γ, an amino acid sequence of the 198th to 462nd positions of mouse retinoic acid receptor α, an amino acid sequence of the 190th to 448th positions of the mouse retinoic acid receptor β, or an amino acid sequence of the 200th to 458th positions of mouse retinoic acid receptor γ.  
     
     
         18 . The transformant according to  claim 6 , wherein the nuclear receptor is a vitamin D 3  receptor.  
     
     
         19 . The transformant according to  claim 6 , wherein the nuclear receptor is a thyroid hormone receptor.  
     
     
         20 . (canceled)  
     
     
         21 . A therapeutic agent for a cancer or an autoimmune disease, comprising a DNA for transforming the transformant of  claim 5  as an active ingredient.  
     
     
         22 . A method for screening an agonist or an antagonist for a intracellular receptor, comprising using the transformant of  claim 5 .  
     
     
         23 . A method for screening an agonist or an antagonist for a PPAR α receptor, a PPAR γ receptor or a PPAR δ receptor, wherein the transformant of  claim 5  is mouse fibroblast L929 or human cancer cell HeLa.