RYANODINE RECEPTOR, ITS GENE AND PRODUCTION OF RYANODINE RECEPTOR

摘要

NEW MATERIAL:A ryanodine receptor composed of 5,037 amino acid residues and containing a specific amino acid sequence. USE:A tranquilizer and a reagent for assay of Ca2+ antagonist. PREPARATION:The objective ryanodine receptor can be produced e.g. by (1) freezing the terminal cisterna of microsome of rabbit skeletal muscle with liquid nitrogen, (2) crushing the frozen product, (3) solubilizing by treating with a homogenizer, (4) centrifuging to collect RNA as a precipitate, (5) separating poly(A) mRNA by conventional process, (6) treating with a reverse transcriptase to obtain a cDNA, (T) treating with a restriction enzyme and bonding to a vector, (8) screening the prepared cDNA bank to select a gene coding a ryanodine receptor, (9) introducing the gene in the form of a DNA fragment into a cloning site downstream of a promoter of an expression vector, (10) inserting into a host and (11) culturing the obtained transformant.