Methods for screening host cells expressing target proteins
AssigneeIndustrial Technology Research Institute
InventorsYen-Ju Lin, Mei-Wei Lin, Min-Yuan Chou
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Abstract
A method for screening host cells expressing a target protein is provided. The method includes the following steps: providing an expression vector, the expression vector including a promoter, a gene encoding a target protein and an FTH1 gene; transfecting the host cells with the expression vector; culturing the host cells in a medium; and adding iron ions to the medium, and screening the surviving host cells to obtain the host cells expressing the target protein. An expression vector and a method for establishing a cell line stably expressing an exogenous recombinant gene are also provided.
Claims10
A method for screening and selecting mammalian host cells expressing a target protein, comprising:
- (a) providing an expression vector comprising a promoter, a gene encoding a target protein and a Ferritin Heavy Chain 1 (FTH1) gene, wherein the FTH1 gene is used as a selection marker;
- (b) transfecting the mammalian host cells with the expression vector;
- (c) culturing the transfected mammalian host cells of step (b) in a culture medium;
- (d) adding 100 μM to 1.5 mM of iron ions and 0.5 μM to 2 mM of a ferroptosis inducer to the culture medium of step (c);
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- (e) screening and selecting the surviving mammalian host cells comprising the expression vector to obtain the mammalian host cells expressing the target protein.
Dependencies are inferred from the claim text.