Hybrid vector having a cytomegalovirus enhancer and myeloproliferative sarcoma virus promoter
View Patent ↗An expression vector capable of expressing high levels of heterologous proteins having a cytomegalovirus (CMV) enhancer 5′ upstream from a myeloproliferative sarcoma virus (MPSV) promoter.
1. The vector pZMP21 as deposited with the ATCC, having the reference number ATCC PTA-5266.
2. The vector of claim 1 further comprises a structural gene such that the gene is operably linked to the cytomegalovirus enhancer and the myeloproliferative sarcoma virus promoter.
3. A mammalian cell transfected with the vector of claim 2 .
4. A method of producing a recombinant protein comprising
a. transfecting a mammalian host cell with the vector of claim 2 ;
b. growing the cells under conditions that selectively propagate those cells that have integrated the vector of claim 1 into its genome;
c. growing the cells of step b) under conditions that cause the recombinant protein to be secreted into the cell medium;
d. isolating the recombinant protein from the cell medium.
5. A method of producing a recombinant protein comprising
a. randomly integrating the vector of claim 2 into the genome of CHO cells;
b. growing the cells in the presence of increasing concentrations of methotrexate;
c. isolating cells from step b) and growing under conditions such that the CHO cells produce the recombinant protein into the culture medium;
d. isolating the recombinant protein from the culture medium.
6. The mammalian cell of claim 3 wherein the cell is a CHO cell.
7. The mammalian cell of claim 6 wherein the CHO cell is of strain DXB 11.
8. The method of claim 4 wherein the transfection occurs by electroporation.
9. The method of claim 4 wherein step b) comprises growing the cells in the presence of methotrexate.
10. The method of claim 5 wherein the CHO cells are of the strain DXB 11.