title: LCR/MEL: a versatile system for high-level expression of heterologous proteins in erythroid cells.
abstract: We have used the human globin locus control region (LCR) to assemble an expression system capable of high-level, integration position-independent expression of heterologous genes and cDNAs in murine erythroleukaemia (MEL) cells. The cDNAs are inserted between the human beta-globin promoter and the second intron of the human beta-globin gene, and this expression cassette is then placed downstream of the LCR and transfected into MEL cells. The cDNAs are expressed at levels similar to those of the murine beta-globin in the induced MEL cells. Heterologous genomic sequences can also be expressed at similar levels when linked to to the LCR and beta-globin promoter. In addition we demonstrate that, after induction of differentiation, MEL cells are capable of secreting heterologous proteins over a prolonged time period, making this system suitable for use in continuous production systems such as hollow fibre bioreactors. The utility of the LCR/MEL cell system is demonstrated by the expression of growth hormone at high levels (greater than 100 mg/l) 7 days after induction. Since the expression levels seen do not depend upon gene amplification and are independent of the integration position of the expression cassette, it is possible to obtain clones with stable high-level expression within 3-4 weeks after transfection.
authors.affiliation: ICI Pharmaceuticals, Biotechnology Department, Macclesfield, Cheshire, UK.
authors.forename: M
authors.initials: M
authors.lastname: Needham
authors.forename: C
authors.initials: C
authors.lastname: Gooding
authors.forename: K
authors.initials: K
authors.lastname: Hudson
authors.forename: M
authors.initials: M
authors.lastname: Antoniou
authors.forename: F
authors.initials: F
authors.lastname: Grosveld
authors.forename: M
authors.initials: M
authors.lastname: Hollis
mesh_terms: D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D005914:Globins / Q000235:genetics*; D013006:Growth Hormone / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D014407:Tumor Cells, Cultured
chemical_list: D013006:Growth Hormone; D005914:Globins