Recombinant protein expression using a hybrid CHEF1 promoter
The invention provides expression vectors and host cells for high-level expression of recombinant proteins. The expression vectors comprise Chinese hamster ovary elongation factor 1-α (CHEF1) transcriptional regulatory DNA elements and a cytomegalovirus (CMV) promoter and/or a human adenovirus tripartite leader (AdTPL) sequence. The invention achieves increased protein expression and better productivity of host cells compared to previously described expression systems.
1. An expression vector comprising Chinese Hamster Elongation Factor-1α (CHEF1) transcriptional regulatory DNA and a cytomegalovirus (CMV) promoter and/or an adenovirus tripartite leader (AdTPL) sequence, wherein the CHEF1 transcriptional regulatory DNA comprises Sequence ID NO: 1.
2. An expression vector comprising Chinese Hamster Elongation Factor-1α (CHEF1) transcriptional regulatory DNA and a cytomegalovirus (CMV) promoter and/or an adenovirus tripartite leader (AdTPL) sequence, wherein the CHEF1 transcriptional regulatory DNA comprises Sequence ID NO: 2.
3. The expression vector of claim 2 , further comprising Sequence ID NO: 3.
4. An expression vector comprising Chinese Hamster Elongation Factor-1α (CHEF1) transcriptional regulatory DNA and a cytomegalovirus (CMV) promoter and/or an adenovirus tripartite leader (AdTPL) sequence, wherein the CHEF1 transcriptional regulatory DNA comprises Sequence ID NO: 3.
5. The expression vector of claim 2 comprising a CMV promoter.
6. The expression vector of claim 2 comprising an AdTPL sequence.
7. The expression vector of claim 2 comprising a CMV promoter and an AdTPL sequence.
8. The expression vector of claim 2 comprising Sequence ID NO: 4.
9. The expression vector of claim 2 comprising Sequence ID NO: 5.
10. The expression vector of claim 2 comprising Sequence ID NO: 6.
11. The expression vector of claim 2 comprising Sequence ID NO: 7.
12. The expression vector of claim 2 , further comprising a selectable marker gene.
13. The expression vector of claim 2 , further comprising a polynucleotide encoding a protein of interest operably linked to the CHEF1 transcriptional regulatory DNA, the CMV promoter and/or the AdTPL sequence.
14. A host cell transformed, transduced or transfected with an expression vector according to claim 2 .
15. The host cell of claim 14 , wherein the host cell is a prokaryotic cell.
16. The host cell of claim 14 , wherein the host cell is a eukaryotic cell.
17. The host cell of claim 16 , wherein the host cell is a hamster cell.
18. The host cell of claim 17 , wherein the host cell is a Chinese Hamster Ovary (CHO) cell.
19. The host cell of claim 16 , wherein the host cell is a non-hamster mammalian cell.
20. The host cell of claim 19 , wherein the host cell is a human cell.