Pigment epithelium-derived factor: characterization, genomic organization and sequence of the PEDF gene

Abstract

Nucleic acids encoding the neurotrophic protein known as pigment epithelium-derived factor (PEDF), a truncated version of PEDF referred to as rPEDF, and equivalent proteins, vectors comprising such nucleic acids, host cells into which such vectors have been introduced, recombinant methods for producing PEDF, rPEDF, and equivalent proteins, the rPEDF protein and equivalent proteins of rPEDF and PEDF-BP, -BX and BA, and the PEDF protein produced by recombinant methods. Effects and uses of these variants on 1) neuronal differentiation (neurotrophic effect) 2) neuron survival (neuronotrophic effect) and 3) glial inhibition (gliastatic effect) are described.

1 . An isolated and purified nucleic acid selected from the group consisting of nucleic acids comprising SEQ ID NO:1, SEQ ID NO:43 and conservatively modified variants thereof.  
     
     
         2 . A protein selected from the group consisting of proteins comprising SEQ ID NO:3 and conservatively modified variants thereof.  
     
     
         3 . An isolated and purified nucleic acid which encodes a protein selected from the group consisting of proteins comprising the sequence of SEQ ID NO:3 and conservatively modified variants thereof.  
     
     
         4 . A vector which comprises a nucleic acid molecule of  claim 1 .  
     
     
         5 . A vector according to  claim 4 , wherein the nucleic acid comprises SEQ ID NO:1.  
     
     
         6 . A vector according to  claim 4 , wherein the nucleic acid comprises SEQ ID NO:43.  
     
     
         7 . A vector which comprises the nucleic acid of  claim 2 .  
     
     
         8 . A vector which comprises the nucleic acid of  claim 3 .  
     
     
         9 . A host cell containing the vector of  claim 5 .  
     
     
         10 . A host cell containing the vector of  claim 6 .  
     
     
         11 . A host cell containing the vector of  claim 7 .  
     
     
         12 . A host cell containing the vector of  claim 8 .  
     
     
         13 . A method of producing the protein encoded by a nucleic acid of SEQ ID NO:1 and conservatively modified variants thereof, said method comprising: 
 expressing said protein in a host cell of  claim 9;  and    recovering said protein from the host cell.    
     
     
         14 . A method of producing a protein comprising SEQ ID NO:2 and conservatively modified variants thereof, said method comprises: 
 expressing said protein in a host cell of  claim 9;  and    recovering said protein from the host cell.    
     
     
         15 . A method of producing a protein comprising SEQ ID NO:3 and conservatively modified variants thereof, said method comprises: 
 expressing said protein in a host cell of  claim 11;  and    recovering said protein from the host cell.    
     
     
         16 . A recombinantly produced PEDF protein, which has been produced in accordance with the method of  claim 14  and which is substantially free from proteins normally associated with a naturally occurring source organism.  
     
     
         17 . An isolated and purified human genomic DNA molecule which encodes a protein having a sequence of SEQ ID NO:2.  
     
     
         18 . The method of  claim 14 , wherein said method further comprises lysing said host cell and then isolating said protein.  
     
     
         19 . The method of  claim 18 , wherein said method further comprises purifying said protein.  
     
     
         20 . The method of  claim 15 , wherein said method further comprises lysing said host cell and then isolating said protein.  
     
     
         21 . The method of  claim 20 , wherein said method further comprises purifying said protein.  
     
     
         22 . The nucleic acid of  claim 1  which comprises the sequence of SEQ ID NO:1.  
     
     
         23 . The vector of  claim 4  comprising a nucleic acid encoding a protein of SEQ ID NO:3.  
     
     
         24 . A recombinantly produced PEDF protein, which has been produced in accordance with the method of  claim 15  and which is substantially free from proteins normally associated with a naturally occurring source organism.  
     
     
         25 . An isolated and purified PEDF protein comprising SEQ ID NO:2 and conservatively modified variants thereof.  
     
     
         26 . A DNA encoding PEDF comprising SEQ ID NO:43.