ASSAY, RECEPTOR PROTEIN NYK, AND LIGAND

摘要

PROBLEM TO BE SOLVED: To establish the large-volume regulation of the extracellular domain of an NYK receptor and a method for detecting receptors and ligand in an assay by bringing receptor protein NYK, its derivative, or its functionally equivalent body into contact with each other under a condition suitable for enabling binding with estimated ligand. SOLUTION: A mutant receptor is cut by BgIII or BamHI and is linked with an FLAGTM linker oligonucleotide to select a recombinant containing extracellular domain arrangement and in-frame FLAGTM arrangement. First, the ability of FLAGTM-fusing protein to come to manifestation is assayed by transient manifestation in a COS cell, and an insert is transferred to a CHO cell manifestation vector through the use of Xbal. A phosphorylated FLAGTM linker oligonucleotide (39-mer) easily linked with a cut vector fragment and the recombinant are accurately detected through the use of an internal CIaI section.