Enhance protein production method
AssigneeBayer Aktiengesellschaft
InventorsMyung-Sam Cho, Van-Mai Tran
Life of this patent
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Abstract
An expression vector comprising a promoter, a coding sequence of a heterologous protein, the coding sequence being operably linked to the promoter, and an intronic sequence downstream of the promoter and upstream of the coding sequence, the intronic sequence comprising two identical donor sites and one acceptor site.
Claims13
An expression vector comprising
- a) a promoter,
- b) a coding sequence of a heterologous protein, the coding sequence being operably linked to the promoter, and
- c) a MIS sequence, the MIS sequence being downstream of the promoter and upstream of the coding sequence.
A DNA molecule comprising the following DNA sequence:
- ##STR1##
A method of increasing stable protein expression of mammalian cell lines comprising the steps of
- a) transfecting a population of mammalian cells with an expression vector comprising an amplifiable marker and a MIS sequence,
- b) growing the cells in the presence of an amplifying agent, and
- c) selecting at least one cell line having high productivity from the cells of step b.
The method of 8, wherein the mammalian cells are selected from the group consisting of human embryonic kidney cells, human B-cells, chinese hamster ovary cells, baby hamster kidney cells, mouse myeloma cells, cells derived from human embryonic kidney cells, cells derived from human B-cells, cells derived from chinese hamster ovary cells, cells derived from baby hamster kidney cells, and cells derived from mouse myeloma cells.
A method of increasing the number of copies of DNA from a vector integrated into a cell's chromosomal DNA, the method comprising
- a) transfecting a population of cells with an expression vector having a promoter, a coding sequence, and a MIS sequence located between the promoter and coding sequence, the expression vector further comprising an amplifiable marker,
- b) growing the cells in the presence of an amplifying agent, and
- c) selecting at least one cell from the cells of step b having an increased number of vector DNA copies integrated into chromosomal DNA.
Dependencies are inferred from the claim text.