Click here to close Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly. We suggest using a current version of Chrome, FireFox, or Safari.
XB-ART-28122
Proc Natl Acad Sci U S A 1987 Jun 01;8412:4332-6.
Show Gene links Show Anatomy links

cDNA cloning of a serotonin 5-HT1C receptor by electrophysiological assays of mRNA-injected Xenopus oocytes.

Lübbert H , Hoffman BJ , Snutch TP , van Dyke T , Levine AJ , Hartig PR , Lester HA , Davidson N .


???displayArticle.abstract???
We describe a strategy for the cloning of neurotransmitter-receptor and ion-channel cDNAs that is based on electrophysiological assays of mRNA-injected Xenopus oocytes. This procedure circumvents the purification of these membrane proteins, which is hindered by their low abundance and their hydrophobic nature. It involves methods for RNA fractionation by high-resolution gel electrophoresis, directional cDNA cloning in a single-stranded vector, and screening of the cDNA library by voltage-clamp measurements of currents induced by serotonin in mRNA-injected oocytes. The applicability of our approach is demonstrated by the isolation of a serotonin receptor cDNA clone from a mouse choroid plexus papilloma. The clone was identified by hybrid-depletion and hybrid-selection procedures. The receptor expressed in oocytes injected with hybrid-selected RNA is fully functional, indicating that it is composed of a single subunit encoded by a 5-kilobase RNA. The pharmacology of the hybrid-selected receptor confirms that we have successfully cloned a serotonin 5-HT1C receptor cDNA.

???displayArticle.pubmedLink??? 3473504
???displayArticle.pmcLink??? PMC305079
???displayArticle.link??? Proc Natl Acad Sci U S A
???displayArticle.grants??? [+]


References [+] :
Bailey, Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. 1976, Pubmed