Human BMP-7 promoter and method for exploring bone-related substance by using the same

Abstract

The present invention provides a method for exploring low molecular weight compounds which regulate positively or negatively the expression of human BMP-7 with reference to a reporter activity by using 5′ upstream region gene containing the human BMP-7 promoter and an animal cell introduced with the vector that has been connected to an appropriate reporter gene. The low molecular weight compounds and their derivatives obtained by the present method have morphogenetic activity or inhibiting activity for bone and cartilage through the expression of the human BMP-7 and are useful as preventive or therapeutic agents for cartilage and bone diseases. Furthermore, the low molecular weight compounds and their derivatives are useful as therapeutic agents for kidney diseases.

What is claimed is:  
     
         1 . A DNA whose nucleotide sequence is represented by the base sequence No. from 1 to 10877 shown in SEQ ID NO. 1 of the Sequence Listing that encodes a human bone morphogenetic protein-7 promoter region, or a fragment thereof.  
     
     
         2 . A method for preparing the DNA shown in SEQ ID NO. 1 of the Sequence Listing comprising the steps of: 
 (1) digestion of a human placenta genomic DNA with a HindIII restriction enzyme,    (2) isolation by agarose gel electrophoresis,    (3) cloning of the isolated DNA fragment digested with HindIII into a lambda phage vector λDASH II treated with the same enzyme,    (4) packaging of said vector into the phage,    (5) establishment of genomic DNA library by infecting  Escherichia coli  with the phage,    (6) screening by PCR, and    (7) subcloning into a plasmid vector.    
     
     
         3 . A recombinant expression vector characterized by intearation of the full length or a part of DNA shown in SEQ ID NO. 1 of the Sequence Listing into a reporter gene.  
     
     
         4 . The method for exploring a bone-related substance characterized by using the recombinant expression vector according to  claim 3 .  
     
     
         5 . The method for exploring a bone-related substance according to  claim 4 , wherein the bone-related substance is osteogenesis inducing substance.  
     
     
         6 . The method for exploring a bone-related substance according to  claim 4 , wherein the bone-related substance is osteogenesis inhibiting substance.  
     
     
         7 . The method for exploring a substance effective at the treatment of a kidney disorder by using a recombinant expression vector according to  claim 3.