A mouse hybrid cell line that supports gene expression from a variety of promoters in amplifiable vectors.
A hybrid cell line was constructed by fusion of mouse L-cells with an NIH3T3 cell line derivative containing a hybrid gene consisting of the mouse immunoglobulin kappa (IgK) variable gene promoter linked to the Escherichia coli gpt gene. Such hybrids grew to a much higher density compared to either of the parental cell lines. The utility of this cell line as a host to express foreign genes was tested by the expression of TGF-beta cDNA using the cytomegalovirus promoter. The vector also contained the human dihydrofolate reductase (DHFR) gene driven by SV40 early promoter, to allow for the amplification of the transfected gene. Initial transformants, selected at 100 nM methotrexate (MTX), were subsequently selected for resistance to a higher concentration of MTX (2 microM). Such clones expressed an increased level of TGF-beta when compared to the initial transformants. Both the initial transformants and the clones with the amplified DHFR gene produced TGF-beta in an acid-activatable precursor form. This mouse hybrid host cell line also allowed the expression of foreign genes cloned in an eukaryotic expression vector with the mouse IgK variable region promoter and human growth hormone as the reporter gene, whereas such vectors did not function in CHO cells. The mouse hybrid cell line was also found to be capable of being used with a broad range of promoters.
Gopal
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Otsuka Pharmaceutical Co., Ltd., Maryland Research Laboratories, Molecular Biology Department, Rockville 20850.
Polte
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Arthur
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Seidman
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D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D002455:Cell Division; D002460:Cell Line; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D013006:Growth Hormone / Q000235:genetics; D006822:Hybrid Cells / Q000502:physiology*; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D007739:L Cells; D051379:Mice; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014162:Transfection; D015291:Transforming Growth Factors / Q000235:genetics
D007145:Immunoglobulin kappa-Chains; D011994:Recombinant Proteins; D015291:Transforming Growth Factors; D013006:Growth Hormone