Expression vectors and cell lines expressing vascular endothelial growth factor D
View Patent ↗This invention relates to expression vectors comprising VEGF-D and its biologically active derivatives, cell lines stably expressing VEGF-D and its biologically active derivatives, and to a method of making a polypeptide using these expression vectors and host cells. The invention also relates to a method for treating and alleviating melanomas or tumors expressing VEGF-D and various diseases.
1. A mammalian cell line transformed or transfected with a promoter operably linked to a VEGF-D nucleic acid, wherein the VEGF-D nucleic acid comprises a nucleotide sequence encoding a continuous portion of SEQ ID NO: 11 having a biological activity of VEGF-D selected from the group consisting of ability to stimulate one or more of endothelial cell proliferation, differentiation, migration, survival or vascular permeability,
wherein the VEGF-D nucleic acid lacks the codons encoding amino acids 203-354 of SEQ ID NO: 11; and
wherein said promoter is capable of promoting expression of the nucleic acid in a mammalian cell.
2. A mammalian cell line transformed or transfected with a promoter operably linked to a VEGF-D nucleic acid, wherein the VEGF-D nucleic acid comprises a nucleotide sequence encoding a continuous portion of SEQ ID NO: 11 having a biological activity of VEGF-D selected from the group consisting of ability to stimulate one or more of endothelial cell proliferation, differentiation, migration, survival or vascular permeability,
wherein the VEGF-D nucleic acid lacks the codons encoding amino acids 203-354 of SEQ ID NO: 11, and
wherein said cell line is the 293-EBNA human embryonal kidney cell line.
3. A mammalian call line transformed or transfected with a promoter operably linked to a VEGF-D nucleic acid, wherein the VEGF-D nucleic acid comprises a nucleotide sequence encoding a continuous portion of SEQ ID NO: 11, wherein the VEGF-D nucleic acid encodes amino acids 2-202 of SEQ ID NO: 11 and wherein the VEGF-D nucleic acid lacks the codons encoding amino acids 203-354 of SEQ ID NO: 11.
4. An expression vector comprising
a nucleic acid comprising a nucleotide sequence encoding a continuous portion of
SEQ ID NO:11 having a biological activity of VEGF-D selected from the group consisting of ability to stimulate one or more of endothelial cell proliferation, differentiation, migration, survival or vascular permeability, wherein the nucleic acid lacks the codons encoding positions 203-354 of SEQ ID NO: 11, and,
a promoter operably linked to the nucleic acid, wherein said promoter is capable of promoting expression of the nucleic acid in a mammalian cell.
5. An isolated nucleic acid comprising a nucleotide sequence encoding a continuous portion of SEQ ID NO:11 having a biological activity of VEGF-D selected from the group consisting of ability to stimulate one or more of endothelial cell proliferation, differentiation, migration, survival or vascular permeability, wherein the nucleic acid lacks the codons encoding amino acids 203-354 of SEQ ID NO: 11.
6. The isolated nucleic acid according to claim 5 , further comprising sequence that encodes an affinity tag attached in-frame to the nucleotide sequence.
7. An expression vector comprising the nucleic acid according to claim 6 .
8. An isolated host cell transformed or transfected with the isolated nucleic acid of claim 5 or the vector according claim 4 .