Expression and cell membrane localization of rat M3 muscarinic acetylcholine receptor produced in Sf 9 insect cells using the baculovirus system

摘要

>A recombinant baculovirus bearing the cDNA coding for the rat muscarinic acetylcholine receptor subtype M3 placed under the control of the Autographa californica nuclear polyhedrosis virus polyhedrin gene promoter, was constructed. Polymerase chain reaction screening was used to identify the recombinant baculovirus. Northern blot analysis of total RNA from insect cells infected with the recombinant baculovirus indicated that the transcripts were abundant. Binding assays carried out with the muscarinic antagonist [3H]quinuclidinyl benzilate indicated that more than 1 × 106 receptors were produced per cell. Immunofluorescence microscopy showed that the receptor is located on the cell surface.