Expression of the human mu opioid receptor in a stable Sf9 cell line.
The cDNA that encodes the human mu opioid receptor (hMOR) has been cloned and expressed in Spodoptera frugiperda (Sf9) cells using a nonlytic vector system. The coding sequence fused to the cleavable glycoprotein signal peptide gp 64, and a C-terminal histidine tag was placed under the transcriptional control of the Orgyia pseudotsugata multicapsid nucleopolyhedrosis virus immediate-early 2 (OpIE2) promoter. Transfected cells were selected using Zeocin resistance and the receptor was constitutively expressed at approximately 12000 receptors per cell. Immunofluorescence images illustrated that more than 75% of the Sf9 cells expressed hMOR at the plasma membrane. This is the first report of the constitutive and heterologous expression of a G protein-coupled receptor in a stably transfected Sf9 cell line, under the control of the OpIE2 promoter.
Kempf
Juliette
J
Département des Récepteurs et Protéines Membranaires, CNRS UPR 9050, Ecole Supérieure de Biotechnologie de Strasbourg, F-67400, Illkirch-Graffenstaden, France.
Snook
Laelie A
LA
Vonesch
Jean Luc
JL
Dahms
Tanya E S
TE
Pattus
Franc
F
Massotte
Dominique
D
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001709:Biotechnology; D002460:Cell Line; D019204:GTP-Binding Proteins / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D011401:Promoter Regions, Genetic / Q000235:genetics; D017450:Receptors, Opioid, mu / Q000235:genetics*; D018411:Spodoptera; D014162:Transfection
D017450:Receptors, Opioid, mu; D019204:GTP-Binding Proteins