[Production of soluble form of human TNF-alpha ligand-binding domain type 1 receptor by expression in Drosophila cells].
5His-tagged human TNFalpha type I receptor (TNFR1) ligand-binding domain was produced in Drosophila cells under control of metallothionein Cu-inducible promoter and purified by Ni-NTA affinity chromatography to homogeneity. TNFR1 gene fragment was cloned by PCR from CD8+ in vitro cultured T-killer normal linage cDNA. In despite of three disulfide bonds, the recombinant protein was correctly folded which was conformed by TNFalpha ligand binding assay in ELISA variant.
Surovtseva
E V
EV
Anikeeva
N V
NV
Sikulev
Iu K
IuK
Shevelev
A B
AB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003300:Copper / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics; D004330:Drosophila / Q000737:chemistry / Q000166:cytology / Q000235:genetics*; D006801:Humans; D008668:Metallothionein / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000187:drug effects; D017510:Protein Folding; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry / Q000276:immunology
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D047888:Receptors, Tumor Necrosis Factor, Type I; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D003300:Copper; D008668:Metallothionein