Internal ribosome entry site of the labial gene for protein expression

Abstract

The invention describes compositions and methods for recombinant protein expression in a wide range of cell types. The compositions comprise an IRES sequence from the Drosophila labial (lab) gene, or a variant or fragment thereof, or alternatively, a homolog of a lab IRES, or a variant or fragment thereof. Methods of using the compositions are also described.

1 .- 5 . (canceled)  
     
     
         6 . A nucleic acid vector for the expression of at least two cistrons comprising: 
 a. a promoter operably linked to a nucleotide sequence comprising at least two cistrons; and    b. at least one nucleotide sequence comprising a homolog of SEQ ID NO. 1, or a variant or fragment thereof, operably linked to at least one of said at least two cistrons, wherein said homolog of SEQ ID NO. 1, or variant or fragment thereof, provides IRES activity.    
     
     
         7 . The nucleic acid vector of  claim 6 , wherein said homolog of SEQ ID NO. 1 comprises SEQ ID NO. 2.  
     
     
         8 . The nucleic acid vector of  claim 6 , wherein at least one of said at least two cistrons comprises a reporter gene.  
     
     
         9 . The nucleic acid vector of  claim 6 , wherein at least one of said at least two cistrons comprises a therapeutic gene.  
     
     
         10 . A biological vector capable of expressing said at least two cistrons comprising the nucleic acid vector of  claim 6 .  
     
     
         11 . The biological vector of  claim 10 , wherein said biological vector is selected from poxvirus, adenovirus, herpesvirus, adeno-associated virus, retrovirus, and baculovirus.  
     
     
         12 .- 14 . (canceled)  
     
     
         15 . A host cell comprising the nucleic acid vector of  claim 6 .  
     
     
         16 . The host cell of  claim 15 , wherein said host cell is a mammalian cell.  
     
     
         17 . (canceled)  
     
     
         18 . A method for expressing at least two cistrons comprising introducing into a host cell a nucleic acid vector comprising the nucleic acid vector of  claim 6 .  
     
     
         19 . The method of  claim 18 , wherein said homolog of SEQ ID NO. 1 comprises SEQ ID NO. 2.  
     
     
         20 .- 26 . (canceled)  
     
     
         27 . A baculovirus transfer vector for the expression of at least two cistrons comprising: 
 a. a polyhedrin promoter operably linked to a nucleotide sequence comprising at least two cistrons; and    b. at least one nucleotide sequence comprising a homolog of SEQ ID NO. 1, or a variant or fragment thereof, operably linked to at least one of said at least two cistrons, wherein said homolog, or variant or fragment thereof provides IRES activity.    
     
     
         28 . The baculovirus transfer vector of  claim 27 , wherein said homolog comprises SEQ ID NO. 2.  
     
     
         29 . The baculovirus transfer vector of  claim 27 , wherein at least one of at least two cistrons comprises a reporter gene.  
     
     
         30 . The baculovirus transfer vector of  claim 27 , wherein at least one of at least two cistrons comprises a therapeutic gene.  
     
     
         31 . A recombinant baculovirus capable of expressing at least two cistrons in a host cell comprising a baculovirus genome comprising: 
 a. a polyhedrin promoter operably linked to a nucleotide sequence comprising at least two cistrons; and    b. at least one nucleotide sequence comprising a homolog of SEQ ID NO. 1, or a variant or fragment thereof operably linked to at least one of said at least two cistrons, wherein said homolog, or variant or fragment thereof, provides IRES activity.    
     
     
         32 . The recombinant baculovirus of  claim 31 , wherein said homolog comprises SEQ ID NO. 2.  
     
     
         33 . A method for producing a recombinant baculovirus capable of expressing at least two cistrons comprising: 
 a. introducing a baculovirus transfer vector of  claim 27  and a baculovirus genomic DNA into a baculovirus host cell so as to effect homologous recombination; and    b. isolating a recombinant baculovirus.    
     
     
         34 . The method of  claim 33 , wherein said recombinant baculovirus is isolated by selecting plaques expressing at least one of said at least two cistrons.  
     
     
         35 . A baculovirus host cell expressing at least two cistrons comprising the recombinant baculovirus of  claim 31 .  
     
     
         36 . A method of treating a patient comprising administering the nucleic acid vector of claim  1  or  6 .  
     
     
         37 . A method of treating a patient comprising administering the biological vector of claim  4  or  10 .  
     
     
         38 . A method of treating a patient comprising: 
 a. excising a cell or tissue from said patient;    b. introducing the nucleic acid vector of claim  1  or  6  into said excised cell or tissue; and    c. reimplanting said cell or tissue into said patient.    
     
     
         39 . A method of treating a patient comprising: 
 a. excising a cell or tissue from said patient;    b. introducing the biological vector of claim  4  or  10  into said excised cell or tissue; and    c. reimplanting said cell or tissue into said patient.