Novel PTH responsive gene

Abstract

This invention relates to a novel PTH gene. The invention further relates to methods of screening, diagnosis and development of therapies for bone related disorders.

1 . An isolated nucleic acid fragment encoding a parathyroid hormone anabolic induced gene in bone (PAIGB) polypeptide selected from the group consisting of: 
 (a) an isolated nucleic acid fragment encoding SEQ ID NO:2,4,6, 8 and 10,    (b) an isolated nucleic acid fragment encoding an amino acid sequence having at least 85% identity with the SEQ ID NO: 2,4,6, 8 and 10,    (c) an isolated nucleic acid molecule that hybridizes with the isolated nucleic acid fragment of (a) under hybridization conditions of 6×SSC (1M NaCl), 45 to 50% formamide, 1% SDS at 37° C., and a wash in 0.5× to 1×SSC at 55 to 60° C.; and,    (d) an isolated nucleic acid fragment that is complementary to (a), (b), or (c).    
     
     
         2 . The isolated nucleic acid fragment of  claim 1  selected from the group consisting of SEQ ID NO:1, 3, 5, 7, and 9.  
     
     
         3 . The isolated nucleic acid fragment of  claim 1  wherein the isolated nucleic acid fragment is DNA.  
     
     
         4 . The isolated nucleic acid fragment of  claim 1  wherein the isolated nucleic acid fragment is RNA.  
     
     
         5 . The isolated nucleic acid fragment of  claim 1  wherein the nucleic acid fragment encodes a polypeptide which is involved in PTH signaling pathway.  
     
     
         6 . The isolated nucleic acid fragment of  claim 1  which when overexpressed induces bone forming activity in bone tissues.  
     
     
         7 . The isolated nucleic acid fragment of  claim 1 , which is overexpressed in response to intermittent PTH administration.  
     
     
         8 - 12 . (canceled)  
     
     
         13 . A chimeric construct comprising the isolated nucleic acid fragment of  claim 1  operatively linked to suitable regulatory sequences.  
     
     
         14 . A host cell transformed with the chimeric construct of  claim 13 .  
     
     
         15 . The host cell of  claim 14  wherein the host cell is selected from the group consisting of an eukaryotic, a prokaryotic cell, and a multicellular organism.  
     
     
         16 . The host cell of  claim 15  wherein the host cell is a mammalian cell.  
     
     
         17 . The host cell of  claim 16  wherein the host cell is a mammalian osteoblast cell.  
     
     
         18 . The host cell of  claim 16  wherein the host cell is selected from the group consisting of COS-7 (monkey kidney), 293 (human kidney), CHO (hamster ovary), HepG2 (human liver), HeLa (human cervical), NIH3T3 (mouse fibroblast), Primary osteoblasts, TE-85 (human osteoblast), MG-63 (human osteoblast), SAOS-2 (human osteoblast), UMR 106 (rat osteoblast), ROS 17/2.8 (rat osteoblast), MC3T3 (mouse osteoblast), and U2OS (human osteoblast)).  
     
     
         19 . The host cell of  claim 15  wherein the host cell is a human osteoblast cell.  
     
     
         20 . The host cell of  claim 15  wherein the host cell is  E. coli.    
     
     
         21 . The host cell of  claim 20  wherein the host cell is selected from the group consisting of DH5alpha, BL21, and DH10B.  
     
     
         22 . The host cell of  claim 15  wherein the host cell is a yeast cell.  
     
     
         23 . The host cell of  claim 22  wherein the host cell is selected from the group consisting of  Schizasaccharomyces, Saccaromyces Cerevisice, Pichia Pastoris,  and  Pichia Methanolic.    
     
     
         24 . The host cell of  claim 15  wherein the host cell is an insect cell.  
     
     
         25 . The host cell of  claim 24  wherein the host cell is selected from the group consisting of SF, SF21 —Spodoptera Frugiperda,  S2 Schneider Cells, and High Five Cells from  Trichoplusia ni  egg.  
     
     
         26 . A vector comprising the isolated nucleic acid fragment of  claim 1 .  
     
     
         27 . The vector of  claim 26 , wherein the vector is a plasmid.  
     
     
         28 . A transformed cell comprising the vector of  claim 26 .  
     
     
         29 . The transformed cell of  claim 28 , wherein the host microorganism is selected from the group consisting of eukaryotic, prokaryotic cell, and multicellular organism.  
     
     
         30 . The transformed cell of  claim 29 , wherein the host microorganism is a mammalian cell.  
     
     
         31 . The host cell of  claim 30  wherein the host cell is a mammalian osteoblast cell.  
     
     
         32 . The transformed cell of  claim 28 , wherein the host microorganism is  E. coli.    
     
     
         33 . The transformed cell of  claim 28 , wherein the host microorganism is a yeast cell.  
     
     
         34 - 37 . (canceled)  
     
     
         38 . A composition for regulating bone-forming activity in a mammal comprising the isolated nucleic acid fragment of  claim 1 .  
     
     
         39 . A composition according to  claim 38 , wherein said parathyroid hormone anabolic induced gene in bone (PAIGB) nucleic acid fragment is from human osteoblast cells.  
     
     
         40 . A composition according to  claim 38 , wherein the bone forming activity is the regulation of bone growth.  
     
     
         41 . A composition according to  claim 38 , wherein the bone forming activity is regulation of bone density.  
     
     
         42 - 88 . (canceled)