title: [Expression of human lymphotoxin gene in CHO-DHFR- cells].
abstract: A recombinant plasmid p91-HuLT was constructed with a human lymphotoxin (HuLT) gene 2.4 kb EcoRI fragment and a mammalian cell expression vector plasmid p91023. The HuLT EcoRI DNA fragment covers entire coding sequence and some 3(1) non-translated region sequence of the gene, p91-HuLT DNA was used to transfect Chinese hamster cell line CHO-DHFR- cells by using calcium phosphate-DNA coprecipitation method, and DHFR+ transfectants were obtained. RNA dot blot hybridization analysis showed that the HuLT mRNA was produced by p91-HuLT in transfectants. The results of MTT dye reduction assay indicated that the DHFR+ cell lines we obtained constitutively synthesize and secrete the HuLT with the cytotoxic activity of at least 200 units per mL of medium.
authors.affiliation: Institute of Genetics, Fudan University, Shanghai.
authors.forename: J T
authors.initials: JT
authors.lastname: Liu
authors.forename: C B
authors.initials: CB
authors.lastname: Li
authors.forename: T H
authors.initials: TH
authors.lastname: Ni
authors.forename: Y M
authors.initials: YM
authors.lastname: Zhou
authors.forename: S Y
authors.initials: SY
authors.lastname: Zhao
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D015870:Gene Expression; D008233:Lymphotoxin-alpha / Q000096:biosynthesis / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D013762:Tetrahydrofolate Dehydrogenase / Q000032:analysis; D014162:Transfection
chemical_list: D008233:Lymphotoxin-alpha; D011994:Recombinant Proteins; D013762:Tetrahydrofolate Dehydrogenase