Transcriptional regulatory element and its use in enhancing the expression of heterologous protein

Abstract

Provided is a polynucleotide the polynucleotide can be used as a WXRE transcriptional regulatory element used to increase the protein expression level of a protein expression system. A protein expression vector or a protein expression systems comprising the above-mentioned WXRE transcriptional regulatory element as well as the use thereof are also provided. The use of the WXRE transcriptional regulatory element can increase the expression level of a heterologous protein greatly with its biological activity unchanged.

1 .- 20 . (canceled) 
     
     
         21 . A vector comprising a heterologous nucleotide sequence and a nucleotide sequence selected from the group consisting of (i) to (ii):
 (i) a sequence having at least 90% sequence identity with any of SEQ ID NOs: 3-9; and   (ii) a sequence having at least 90% sequence identity with a reverse complementary sequence of any of SEQ ID NOs: 3-9.   
     
     
         22 . The vector of  claim 21 , further comprising a nucleotide sequence selected from the group consisting of (iii) to (iv):
 (iii) a sequence having at least 80% sequence identity with SEQ ID NO: 13; and   (iv) a sequence having at least 80% sequence identity with a reverse complementary sequence of SEQ ID NO: 13   
     
     
         23 . The vector of  claim 21 , further comprising a CMV promoter. 
     
     
         24 . The vector of  claim 23 , wherein the CMV promoter comprises a nucleotide sequence having at least 80% sequence identity with SEQ ID NO: 16. 
     
     
         25 . The vector of  claim 23 , wherein the CMV promoter comprises a nucleotide sequence that is identical to SEQ ID NO: 16. 
     
     
         26 . The vector of  claim 22 , wherein the nucleotide sequence selected from the group consisting of (iii) to (iv) has at least 90% sequence identity with SEQ ID NO: 13. 
     
     
         27 . The vector of  claim 22 , wherein the nucleotide sequence selected from the group consisting of (iii) to (iv) is identical to SEQ ID NO: 13. 
     
     
         28 . The vector of  claim 21 , wherein the nucleotide sequence selected from the group consisting of (i) to (ii) has at least 90% sequence identity to SEQ ID NO: 3. 
     
     
         29 . The vector of  claim 21 , wherein the nucleotide sequence selected from the group consisting of (i) to (ii) has at least 90% sequence identity to SEQ ID NO: 4. 
     
     
         30 . The vector of  claim 21 , wherein the nucleotide sequence selected from the group consisting of (i) to (ii) has at least 90% sequence identity to SEQ ID NO: 5. 
     
     
         31 . The vector of  claim 21 , wherein the nucleotide sequence selected from the group consisting of (i) to (ii) has at least 90% sequence identity to SEQ ID NO: 6. 
     
     
         32 . The vector of  claim 21 , wherein the nucleotide sequence selected from the group consisting of (i) to (ii) has at least 90% sequence identity to SEQ ID NO: 7. 
     
     
         33 . The vector of  claim 21 , wherein the nucleotide sequence selected from the group consisting of (i) to (ii) has at least 90% sequence identity to SEQ ID NO: 8. 
     
     
         34 . The vector of  claim 21 , wherein the nucleotide sequence selected from the group consisting of (i) to (ii) has at least 90% sequence identity to SEQ ID NO: 9. 
     
     
         35 . The vector of  claim 21 , wherein the heterologous nucleotide sequence encodes one or more proteins, wherein the one or more proteins are selected from a group consisting of an antibody, a fusion protein, an enzyme, a soluble protein, a membrane protein, a structural protein, a ribosome protein, a zymogen, a cell surface receptor protein, a transcriptional regulatory protein, a translational regulatory protein, a chromatin protein, a hormone, a cell cycle regulatory protein, a G protein, a neuroactive peptide, an immunomodulatory protein, a blood component protein, an ion gate protein, a heat shock protein, a dihydrofolate reductase, an antibiotic resistance protein, and a fragment thereof. 
     
     
         36 . A host cell comprising the vector of  claim 21 . 
     
     
         37 . The host cell of  claim 36 , wherein the host cell is a Chinese hamster ovary (CHO) cell. 
     
     
         38 . A method of preparing a host cell that stably expresses a protein, the method comprising: inserting into a host cell the vector of  claim 21 . 
     
     
         39 . The method of  claim 38 , wherein the host cell is a Chinese hamster ovary (CHO) cell. 
     
     
         40 . A method of preparing a protein, comprising: culturing the host cell of  claim 36  under conditions for production of the protein.