High level expression of the IL-1 receptor related T1 receptor in insect cells.
Baculovirus-dependent expression of membrane-associated and secreted, heavily modified proteins in insect cells often results in low yields. To optimize expression of T1, a heavily glycosylated receptor related to Interleukin-1 receptor type I, deletion mutants comprising either the heavily glycosylated extracellular domain or the isolated transmembrane and cytoplasmic portions of the T1 receptor were expressed from recombinant baculoviruses. As shown here, the use of a Baculovirus-derived leader sequence (GP67) in combination with an insect cell line with increased secretory capacity resulted in yields of several mg per liter (10(9) cells) of culture of both purified secretory T1 glycoprotein and membrane-associated T1 receptor.
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D001483:Base Sequence; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D002846:Chromatography, Affinity; D017931:DNA Primers; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009842:Oligopeptides; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D021382:Protein Sorting Signals / Q000235:genetics; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification; D017472:Receptors, Interleukin-1 / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D017384:Sequence Deletion; D018411:Spodoptera; D014162:Transfection
D017931:DNA Primers; D008565:Membrane Proteins; D009842:Oligopeptides; D021382:Protein Sorting Signals; D011506:Proteins; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins