High level stable expression of pharmacologically active human M1-M5 muscarinic receptor subtypes in mammalian cells.
cDNAs encoding for five mAChR subtypes (M1-M5) were cloned under different promoters in various eukaryotic vectors and each subtype was expressed in different mammalian cell lines. CHO-K1 cell line was the best for generating stable cell lines expressing muscarinic receptors. Immunofluorescence and flow cytometry revealed that expression of M1-M5 was primarily localized on the cell membrane. Western blotting and radio-ligand binding studies revealed that expression of each receptor was stable at higher passages.
Khattar
Sunil K
SK
Department of Biotechnology, Ranbaxy Research Laboratories, Udyog Vihar Industrial Area, Gurgaon, Haryana, India. sunil.khattar@ranbaxy.com
Bora
Roop Singh
RS
Priyadarsiny
Priyanka
P
Gupta
Dikshi
D
Khanna
Alka
A
Narayanan
K Lakshmi
KL
Babu
Venkatesh
V
Chugh
Anita
A
Saini
Kulvinder Singh
KS
D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D011869:Radioligand Assay; D043643:Receptor, Muscarinic M1 / Q000096:biosynthesis* / Q000235:genetics; D043642:Receptor, Muscarinic M5 / Q000096:biosynthesis* / Q000235:genetics
D043643:Receptor, Muscarinic M1; D043642:Receptor, Muscarinic M5