Novel beta-actin and rps21 promoters and uses thereof

Abstract

The invention relates to isolation of novel β-actin and ribosomal protein S21 (rpS21) promoters and uses thereof. In particular, this invention features nucleotide sequences for rodent β-actin promoters including, hamster, rat, and mouse, and hamster rpS21 promoter.

1 . An isolated β-actin promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1, or a variant thereof having promoter activity, wherein the variant is a nucleotide sequence having at least 95% identity to the nucleotide sequence set forth in SEQ ID NO: 1 over the entire length of that reference sequence and wherein the variant is the same length as the nucleotide sequence set forth in SEQ ID NO: 1 or shorter. 
     
     
         2 - 36 . (canceled) 
     
     
         37 . A vector comprising the promoter of  claim 1 . 
     
     
         38 . The vector of  claim 37 , wherein the promoter is operably linked to a heterologous nucleic acid. 
     
     
         39 . The vector of  claim 38 , wherein the heterologous nucleic acid encodes a therapeutic protein. 
     
     
         40 . The vector of  claim 39 , wherein the therapeutic protein is chosen from acid sphingomyelinase, α-glucosidase, and tissue plasminogen activator. 
     
     
         41 . A host cell transfected with the vector of  claim 39 . 
     
     
         42 . The host cell of  claim 41 , wherein the cell is a CHO cell. 
     
     
         43 . The isolated rodent promoter of  claim 1 , wherein the variant is at least 1250 nucleotides in length. 
     
     
         44 . A vector comprising the promoter of  claim 43 . 
     
     
         45 . The vector of  claim 44 , wherein the promoter is operably linked to a heterologous nucleic acid. 
     
     
         46 . The vector of  claim 45 , wherein the heterologous nucleic acid encodes a therapeutic protein. 
     
     
         47 . The vector of  claim 46 , wherein the therapeutic protein is chosen from acid sphingomyelinase, α-glucosidase, and tissue plasminogen activator. 
     
     
         48 . A host cell transfected with the vector of  claim 45 . 
     
     
         49 . The host cell of  claim 48 , wherein the cell is a CHO cell. 
     
     
         50 . A method of producing a protein, comprising:
 (a) culturing a cell transfected with a vector comprising an isolated β-actin promoter operably linked to a heterologous nucleic acid encoding a protein, wherein the isolated β-actin promoter comprises the nucleotide sequence set forth in SEQ ID NO: 1, or a variant thereof having promoter activity, wherein the variant is a nucleotide sequence having at least 95% identity to the nucleotide sequence set forth in SEQ ID NO: 1 over the entire length of that reference sequence and wherein the variant is the same length as the nucleotide sequence set forth in SEQ ID NO: 1 or shorter; and   (b) recovering the protein.   
     
     
         51 . The method of  claim 50 , wherein the protein is an antibody. 
     
     
         52 . The method of  claim 51 , wherein the antibody binds a TGF-β family member. 
     
     
         53 . The method of  claim 50 , wherein the protein is a therapeutic protein. 
     
     
         54 . The method of  claim 53 , wherein the therapeutic protein is chosen from acid sphingomyelinase, α-glucosidase, and tissue plasminogen activator. 
     
     
         55 . A method of producing a protein, comprising:
 (a) culturing a cell transfected with a vector comprising an isolated β-actin promoter operably linked to a heterologous nucleic acid encoding a protein, wherein the isolated β-actin promoter comprises the nucleotide sequence set forth in SEQ ID NO: 1, or a variant thereof having promoter activity, wherein the variant is a nucleotide sequence having at least 95% identity to the nucleotide sequence set forth in SEQ ID NO: 1 over the entire length of that reference sequence and wherein the variant is the same length as the nucleotide sequence set forth in SEQ ID NO: 1 or shorter, provided the variant is at least 1250 nucleotides in length; and   (b) recovering the protein.   
     
     
         56 . The method of  claim 55 , wherein the protein is an antibody. 
     
     
         57 . The method of  claim 56 , wherein the antibody binds a TGF-β family member. 
     
     
         58 . The method of  claim 55 , wherein the protein is a therapeutic protein. 
     
     
         59 . The method of  claim 58 , wherein the therapeutic protein is chosen from acid sphingomyelinase, α-glucosidase, and tissue plasminogen activator.